01 imaging systems pdf thorlabs by Axxis ecuador

Confocal Microscopes (See Pages 54 - 65)

Imaging Systems

Imaging Components

Imaging Cytometry

Electrophysiology

Rigid Stands (See Pages 296 - 300)

1 - 137

138 - 247

248 - 267

268 - 305

Scientific CCD Cameras

306 - 327

Optogenetics

328 - 351

Workstations

352 - 383

CCD Cameras (See Pages 306 - 327)

Accessories

Introduction Bergamo II Series Multiphoton Imaging System (See Pages 2 - 35)

Indices

384 - 567

A1 - A18

586 - 604

Table of Contents

A Bit About Us

Thorlabs, founded in 1989 and still headquartered in Newton, NJ, started with a vision of making optical components readily available. The business model focused on quickly delivering product into the hands of our customers, thus eliminating time constraints. Thorlabs built upon this simple concept of speed by expanding in-house manufacturing capabilities to ensure that fair-priced, quality products could be kept in stock for sameday shipment, in addition to ensuring responsive customer support and a low-friction interface. Now in our 25th year of business, Thorlabs employs over 1200 people, has offices in nine countries across four continents, and manufactures over 90% of the more than 15,000 products in our portfolio. This portfolio, which grew from a handful of photonics tools staples, now also encompasses a variety of imaging components and equipment. Through a number of acquisitions, and by building our manufacturing capabilities, we have expanded our competencies into the Life Science systems market. In 2008, Thorlabs made the strategic growth decision to start a business unit in Sterling, VA, known as Thorlabs Imaging Systems, that would be dedicated to designing and manufacturing imaging components and systems for the life science market. Soon after, the team working on our confocal imaging scan head relocated from NJ to VA, allowing all of Thorlabsâ&#x20AC;&#x2122; laser imaging scientists to work under one roof and mutually benefit from each otherâ&#x20AC;&#x2122;s expertise. With customer feedback and additional internal R&D efforts, we quickly progressed a prototype to a fully commercialized confocal scan head that could be added to a standard inverted microscope to enable confocal imaging. Today, the Thorlabs Imaging Systems team consists of over 40 professionals encompassing specialties in mechanical, electrical, and optical engineering, as well as software, production, and manufacturing. Together, they are tasked with developing sophisticated imaging systems for the life sciences market. Keeping with the Thorlabs philosophy of vertically integrated manufacturing and functioning as a single cohesive team, our products benefit from the highest levels of quality control. In addition, our service philosophy allows us to adapt and respond to rapidly changing research environments. If we do not directly possess the necessary manufacturing capabilities in our Virginia facility, we are able to leverage the in-house capabilities of other Thorlabs divisions and our large network of manufacturing facilities located throughout the world. This unrivaled level of access and integration allows the team to rapidly bring to market intuitive, user-friendly confocal and multiphoton microscopy solutions. As a further testament to our growth and dedication in the life science research community, since 2011, Thorlabs has added the Burleigh family of micromanipulators, Thorlabs Scientific Imaging (formerly DVC Camera Corporation), and most recently, the CompuCyte family of imaging cytometers to our product offerings. Our success is predicated on close interactions with our customers and a deeply vested interest in creating the most advanced imaging systems.

Thorlabs Imaging Systems Team

Imaging Systems Selection Guide

Bergamo Series Microscope

ThorImageLS Software

Pages 2 - 35

Pages 66 - 69

Cerna Series Microscope

OCT Systems

Pages 36 - 49

Pages 70 - 115

Essentials Kit

TDI Digital Microscopy

Pages 50 - 53

Pages 116 - 123

Confocal Microscopes

Optical Tweezers

Pages 54 - 65

Pages 124 - 137

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes

Overview: Bergamo II Series Multiphoton Microscopes

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy

A User Looks Through the Trinoculars to Locate a Region of Interest for Laser Scanning

Optical Tweezers

The Bergamo II Series is Thorlabs’ platform for multiphoton microscopy. Taking advantage of close collaborations with our colleagues in the field, decades of cumulative industry experience, and our extensive manufacturing background, we have engineered a family of modules that enable us to configure each individual microscope to your exact experimental requirements. Our versatile microscopes fully support techniques that require laser scanning, widefield imaging, and transmitted light imaging modalities.

Options at a Glance n Scan

Paths •P rimary Galvo-Resonant and Secondary Galvo-Galvo • Primary Galvo-Resonant • Primary Galvo-Galvo • S uper Broadband Correction Range of 400 - 1100 nm or 600 - 1400 nm n Motion Control for Microscope Body • Rotation About the Focus and XYZ Translation • XYZ Translation • Z Translation n PMT Detection Modules •E xtended FOV, 4 PMTs, Backward Direction •E xtended FOV, 2 PMTs, Backward Direction • Full FOV, 2 PMTs, Backward Direction • Full FOV, 2 PMTs, Forward Direction 2

n Widefield

Viewing • Scientific Camera • Nikon Trinoculars • Thorlabs Single-Cube Epi-Illuminator • Nikon Epi-Illuminator for 6 Filter Cubes •H igh-Power LED or Liquid Light Guide Source n Transmitted Light Imaging • Dodt Contrast • Laser-Scanned Dodt Contrast • Differential Interference Contrast (DIC) • Air or High-NA Oil Immersion Condenser from Nikon n Motion Control Accessories • Piezo Objective Holder • XY Platform • Z-Axis Piezo Stage

Imaging Systems

Overview: Bergamo II Series Multiphoton Microscopes

Bergamo Series Microscopes +

Configure Your Microscope to Your Experiment

The most critical choices in configuring your Bergamo II microscope are the motion control options for the microscope body, the number and type of laser scanning paths, and the number of PMT detection channels. These options are indicated with icons placed next to the systems shown on the following pages. In each of these categories, we supply several options in recognition of your unique experimental requirements.

Cerna Series + Microscopes

R G

R+G R + G Essentials Kit R R+G

2X FFV 2X EFV 4X EFV Confocal Microscopes 2X FFV 2X EFV 4X EFV 2X FFV 2XThorImageLS EFV 4X EFV Software

OCT Systems TDI Digital Microscopy Optical Tweezers

This non-rotating Bergamo II microscope body features Z-axis motorization, an XY platform, a galvoresonant scan path for imaging at up to 400 fps, and two PMT detection channels with extended-fieldof-view collection optics. Non-rotating bodies support our user-installable Dodt contrast and DIC transmitted light imaging modules.

This rotating Bergamo II microscope body features a five-axis motorized body (X, Y, Z, θ, and Elevation), dual scan paths for photoactivation and functional imaging, and four PMT detection channels with extended-fieldof-view collection optics. These choices enable highperformance in vivo imaging.

+ + +

R+G R R+G R R+G

2X EFV 4X EFV FV 2X EFV 4X EFV

LEGEND

2X EFV

4X EFV

+ On the following pages, there are icons next to each system that indicate the key modules chosen for that configuration. Please consult pages 12 – 15 and 18 – 19 for details on these + G R R+G modules.

BODY

SCAN PATHS +

+ Z

R+G

+ +

G +

G G

2X FFV

R+G

XYZ

4X EFV

2X FFV

2X EFV

4X EFV

PMT DETECTION

R+G

2X FFV

2X EFV

4X2 EFV PMTs, Full FOV

G R R+G 2X FFV 2X EFV R + Primary G Galvo-Galvo

4X EFV

2 PMTs, Extended FOV

Primary Galvo-Resonant

R + G 2X FFV 2X EFV 4X EFV XYZ + θ and Primary Galvo-Resonant and 2X FFV 2X EFV G R R+G Elevation Secondary Galvo-Galvo 2X FFV 2X EFV 4X EFV R+G 2X EFV

4X EFV

4 PMTs, Extended FOV

Imaging Systems Bergamo Series Microscopes

Overview: Bergamo II Series Multiphoton Microscopes

Cerna Series Microscopes

Essentials Kit Confocal Microscopes

ThorImageLS Software

2X FFV

OCT Systems TDI Digital Microscopy Optical Tweezers

Rotating Microscopes: The Flexibility to Bring the Objective to the Region of Interest

Like the original Bergamo microscope, the R + G II is available with a five-axis motorized Bergamo body (X, Y, Z, θ, and Elevation) that translates the entire optical path, rotates about a fixed point in 2X EFV 4X EFV space, and creates a large volume underneath the objective for complex experimental apparatuses. Bergamo II systems with rotation can be equipped with a -5° to +95° range (for single and dual scan path configurations) or a symmetric -50° to +50° range (for single scan path configurations only). Since

the microscope moves the entire optical system, it is not necessary to reposition the sample, adjust the focus, or re-locate the region of interest after the objective is rotated. An integrated elevator coarsely moves the objective over a 5" (12.7 cm) range in the Z direction, supporting experimental setups and apparatuses of varying sizes. Motorizing the optical system not only simplifies experiments for the user but also helps ensure that the designed optical performance of your microscope and your objectives is achieved. Our articulating periscope, detailed more on page 13, is a key feature of our rotating design. -5° to +95° Rotation

Dual-scan-path systems, like the one shown here, rotate from -5° to +95°. Single-scan-path systems can be factory-configured for -5° to +95° or -50° to +50°.

Imaging Systems

Overview: Bergamo II Series Multiphoton Microscopes

Non-Rotating Microscopes: One System for In Vitro and In Vivo Experiments +

The Bergamo II design makes substantial

G G R R RtoR+the +GGoptical performance of the improvements original Bergamo microscope. In addition to a 20 mm field of view that is 1.6X larger than the 2X2XFFV FFV 2X2XEFV EFV 4X4XEFV EFV original design and super broadband scan optics corrected for either 400 - 1100 nm or 600 - 1400 nm, we created transmitted light modules that enable Dodt contrast, laser-scanned Dodt contrast, and differential interference contrast (DIC). Engineered for non-rotating Bergamo II bodies, these modules are compatible with air or high-NA oil immersion condensers from Nikon.

Non-rotating Bergamo II systems are designed around Thorlabsâ&#x20AC;&#x2122; 95 mm construction rails (see www.thorlabs.com for details). We chose these rails because they offer stable long-term support, excellent vibrational damping, and a linear dovetail mounting surface. This dovetail allows us to manufacture userinstallable and removable transmitted light modules that take less than 5 minutes to remove or install. In order to perform in vitro experiments, simply attach the modules to the rail, using a balldriver to tighten a few setscrews in place. To make room underneath the objective for in vivo needs, loosen the setscrews and remove the modules. The linear dovetail centers the modules along the optical path so that the user can avoid alignment issues.

Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

The linear dovetail also allows a user to reposition the scan paths vertically along the spine of the microscope, allowing correct placement between various objectives and condensers. Designed with our non-rotating microscope bodies in mind, our rigid stand sample holders (see pages 296 - 300) offer a slim profile that conserves space for both in vitro and in vivo experiments.

Imaging Systems Bergamo Series Microscopes

Rotating Systems

Cerna Series Microscopes

Rotating Entry-Level In Vivo Microscope

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

G G

Configuration B209

With dual detection channels and full-field-of-view collection optics, this configuration is functionally equivalent to the original Bergamo microscope. The single scan path is configured with Thorlabs’ 8 kHz galvo-resonant scanner, which provides a video frame rate of 30 fps at 512 x 512 pixels and a maximum frame rate of 400 fps at 512 x 32 pixels.

from Various Angles Without Needing to Reposition the Sample G R R +2XGFFV 2X EFV 4X EFV n Large Approach Angles Around Objective n Galvo-Resonant Scanner Provides 30 fps at FFV and 2X400 EFVfps at 4X512 EFVx 32 Pixels 512 x 5122X Pixels

Like all rotating Bergamo II microscopes, it can be equipped with a scientific camera to visualize the field of view and locate fiducial markers. Widefield imaging is augmented by our single-cube epi-illuminator, which can be outfitted with one of Thorlabs’ highpower LEDs (see pages 484 - 508) or an industrystandard liquid light guide (see page 511).

Widefield Imaging • Scientific Camera • Thorlabs’ Epi-Illuminator with High-Power LED

• 2 Channels • Full-Field-of-View Collection Optics • Easy-to-Reach Filters

Scan Path Elevator Base • 5" (12.7 cm) of Coarse Z Travel

• Primary Galvo-Resonant • Optics for 600 - 1400 nm

XYZ and Ѳ Motion • -5° to +95° Rotation • 2" Travel in X and Y • 1" Travel in Z

Articulated Periscope • Maintains Optimal Beam Alignment as Microscope Rotates

Motion Controller

• Move Microscope in X, Y, Z, and Ѳ • Variable Speed Control • Touchscreen for Position Readout and Retrieval

Sample Holder • Rigid Stand with Platform

R+

2X R FFV + G 2X EFV

n Image

PMT Detection Module

Imaging Systems

Rotating Systems Rotating Premium In Vivo Microscope This model is our most capable and versatile Bergamo II microscope with rotation. In order to enable targeted photostimulation, we added a secondary galvo-galvo scan path that complements the primary galvo-resonant path. The separately controlled secondary beam stimulates a specific region within the field of view, then the galvo-resonant scan path automatically performs fast functional imaging at up to 400 fps (depending on the pixel dimensions of the desired image). The galvo-galvo scan path may also be used independently from the galvo-resonant scan path. Galvo-galvo scanners can be targeted at specific regions within the field of view to minimize photo exposure to non-targeted areas. They also permit long pixel dwell times (range: 0.4 to 20 µs), helping to capture weak signals through integration.

G 2X FFV R Configuration B248 +

R+G

2X R EFV + G 4X EFV

n Extended-Field-of-View

Collection Optics Harvest Signal Photons Scattered by Thick G R2X FFVR + 2X G EFV 4X EFV In Vivo Preparations n Four PMTs Provide Simultaneous Detection of Four 2X Fluorophores FFV 2X EFV 4X EFV n Secondary Galvo-Galvo Scan Path for PhotoUncaging, Targeted Illumination, and Imaging

Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Widefield Imaging • Scientific Camera • Thorlabs’ Epi-Illuminator with High-Power LED

Dual Scan Paths • Primary Galvo-Resonant for 600 - 1400 nm • Secondary Galvo-Galvo for 400 - 1100 nm

PMT Detection Module • 4 Channels • Extended-Field-of-View Collection Optics • Easy-to-Reach Filters

See a 360º Video of this Configuration at thorlabs.com

Articulated Periscope • Maintains Optimal Beam Alignment as Microscope Rotates

Elevator Base • 5 " (12.7 cm) of Coarse Z Travel

Sample Holder • Rigid Stand with Platform

Imaging Systems Bergamo Series Microscopes

Non-Rotating Systems

Cerna Series Microscopes

Entry-Level In Vitro and In Vivo Microscope with Z Motion

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

This Bergamo II configuration is outfitted with two detection channels and extended-field-of-view collection optics, offering high detection efficiency for both thin in vitro samples and thicker in vivo preparations. Compared to our full-field-of-view optics, these optics provide a larger collection angle that captures more of the signal photons that are scattered by thick tissue.

Configuration B206

G 2X FFV R

R+G

2XR EFV + +G 4X EFV

n 1"

Travel Along the Z Direction with 0.1 µm Resolution 2XGFFV R2X EFVR +4X GEFV n Extended-Field-of-View Collection Optics Gather Signals Generated Deep In Vivo and Improve Signal-to-Noise of In 2X Vitro FFV Samples 2X EFV 4X EFV n Piezo Objective Holder for High-Speed Z-Stack Acquisitions

A motorized, software-controlled dichroic mover makes it possible to switch between laser scanning and widefield imaging modalities remotely from the computer interface.

Widefield Imaging PMT Detection Module • 2 Channels • Extended-Field-of-View Collection Optics • Easy-to-Reach Filters

• Scientific Camera • Thorlabs’ Epi-Illuminator with High-Power LED

Scan Path • Primary Galvo-Resonant • Optics for 600 - 1400 nm

Motorized Dichroic Mover • Computer Controlled

Z Motion • 1" Travel

Piezo Objective Holder • For Fast Z-Stacks

Sample Holder • Rigid Stand with Platform

Microscope Controller • Move Microscope in Z • Variable Speed Control

Imaging Systems

Non-Rotating Systems Mid-Level In Vitro and In Vivo Microscope with Z Motion and XY Platform

Configuration B2322X FFV

To capitalize on our extended-field-of-view collection optics’ design, we engineered a detection module capable of detecting four fluorophores simultaneously. The 4-channel detection module offers the same easy access to optics as the 2-channel version on the previous page, making it simple to exchange emission filters and dichroics so that the microscope can detect different fluorescent tags.

R+G 2X EFV G

4XREFV+

R+G

n XY

To supplement the 1" of Z travel provided by the microscope body, our XY platform has been incorporated into this system. By recessing the 95 mm construction rail that serves as the microscope’s support structure, we are able to provide access to the objective from the front, sides, and even behind.

Bergamo Series Microscopes Cerna Series Microscopes Essentials

Platform Underneath the Objective Moves Kit Sample and Setup as One Confocal G 2XRFFV R2X+ EFV G 4X EFV n Primary Galvo-Galvo Scan Path Provides Microscopes Consistent Pixel Dwell Times Across the ThorImageLS Software Entire Field of View 2X FFV 2X EFV 4X EFV n Four Detection Channels with ExtendedOCT Systems Field-of-View Collection Optics Capture TDI Digital Signals Scattered Through Thick In Vivo Microscopy Preparations

PMT Detection Module • 4 Channels • Extended-Field-of-View Collection Optics • Easy-to-Reach Filters

Optical Tweezers

See a 360º Video of this Configuration at thorlabs.com

Widefield Imaging • Scientific Camera • Thorlabs’ Epi-Illuminator with High-Power LED

Scan Path • Primary Galvo-Galvo • Optics for 600 - 1400 nm

Z Motion • 1" Travel

Sample Holder • XY Platform • 2" Travel in X and Y

Microscope Controller • Move Microscope in Z • Control Position of XY Platform • Variable Speed Control

Imaging Systems Bergamo Series Microscopes

Non-Rotating Systems

Cerna Series Microscopes

Entry-Level In Vitro Microscope with XYZ Motion

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Configuration B203

This Bergamo II system is equipped with three independent, high-resolution motors for hands-free positioning of the objective in X, Y, and Z (X and Y: 2" travel and 0.5 µm resolution; Z: 1" travel and 0.1 µm resolution). It is complemented by a sample holder with our Z-axis piezo stage (detailed on pages 174 - 176), which accepts multiwell plates, multiple slides, petri dishes, or even a small breadboard. The piezo stage allows these accessories to be positioned beneath the objective with 25 nm resolution in Z. This configuration is also equipped with a Nikon epi-illuminator for widefield imaging. Available on non-rotating Bergamo II microscope bodies with full-

2XR FFV + G 2X EFV

n Nikon-Designed

Epi-Illuminator Accepts Liquid Light Guides and up to 6 Filter Cubes G 2XRFFV R2X+ EFV G 4X EFV n Piezo Stage on Rigid Stand Translates Samples with 25 nm Resolution in Z 2X FFV Optics 2X EFVMatch 4X EFV n Full-Field-of-View Collection the Objective’s Back Aperture for Efficient In Vitro Signal Collection field-of-view collection optics, this Nikon-engineered module uses an industry-standard liquid light guide for illumination (see page 511 for our selection) and holds up to six dichroics at once.

See a 360º Video of this Configuration at thorlabs.com

Widefield Imaging

PMT Detection Module • 2 Channels • Full-Field-of-View Collection Optics • Easy-to-Reach Filters

• Nikon Epi-Illuminator for 6 Filter Cubes with Liquid Light Guide • Scientific Camera • Nikon Trinoculars

Scan Path • Primary Galvo-Resonant • Optics for 600 - 1400 nm

XYZ Motion • 2" Travel in X and Y • 1" Travel in Z

Sample Holder • Rigid Stand with Z-Axis Piezo Stage

Controller • Move Microscope in X, Y, and Z • Variable Speed Control •T ouchscreen for Position Readout and Retrieval

R+

Imaging Systems

Non-Rotating Systems Premium In Vitro and In Vivo Microscope with XYZ Motion This configuration incorporates four detection channels with our extended-field-of-view collection optics for deep imaging in vivo and is equipped with dual galvo-resonant and galvo-galvo scan paths for high-frame-rate functional imaging, targeted laser exposure, and photostimulation/ uncaging experiments. It is also outfitted with our Dodt contrast transmitted light module and a Nikon condenser with 1" of motorized travel in Z, highlighting the versatility of our modular microscope construction. Our transmitted light modules can be installed or removed by the user on any non-rotating Bergamo II configuration in just a few minutes, making it

G 2X FFV R Configuration B264

R+G

2X R +EFV G +

Bergamo Series Microscopes Cerna Series Microscopes

4X EFV

Essentials Kit

n Dual

Scan Paths for Photoactivation and Functional Imaging at up to 400 fps 2XGFFV R 2X EFVR +4X GEFV n Can Be Converted by User from In Vitro to In Vivo in Less than 5 Minutes 2X FFV 2X EFV Scan 4X Path EFV n Fiber-Coupled Laser in Secondary for Single-Photon Fluorescence

Confocal Microscopes ThorImageLS Software OCT Systems

exceptionally easy to switch between in vitro and in vivo imaging modalities. Thorlabs’ rigid stand sample holders (see pages 296 - 300) are the ideal match for systems that incorporate these transmitted light modules. Each has a narrow profile that allows slides to rotate easily into and out of the optical path.

TDI Digital Microscopy Optical Tweezers

See a 360º Video of this Configuration at thorlabs.com

Widefield Imaging • Scientific Camera • Thorlabs’ Epi-Illuminator with HPLS Liquid Light Guide

Dual Scan Paths PMT Detection Module • 4 Channels • Extended-Field-of-View Collection Optics • Easy-to-Reach Filters

Fiber-Coupled Laser • I ndependent Laser for Secondary Scan Path

• Primary Galvo-Resonant for 600 - 1400 nm • Secondary Galvo-Galvo for 400 - 1100 nm

Dodt Contrast Module • Installed or Removed by User in Less than 5 Minutes

Condenser Controller • Move Condenser in Z • 1" Travel

Controller • Move Microscope in X, Y, and Z • Variable Speed Control •T ouchscreen for Position Readout and Retrieval

Imaging Systems Bergamo Series Microscopes

Bergamo II Modules in Detail

Cerna Series Microscopes

Galvo-Resonant and/or Galvo-Galvo G GR Scan Paths

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

G RR or +G RR+ G R + G

The number and type of scanning paths 2X FFV 2X FFV 2X 2X EFV 2X FFV EFV 4X 2X EFV 4X EFV EFV 4X EFV determine the imaging modalities that the microscope supports. Each Bergamo II microscope can be configured with galvoresonant and/or galvo-galvo scan paths. Galvo-resonant scanners permit the highest refresh rates (up to 400 fps at 512 x 32 pixels) and are frequently used with Pockels cells (see page 149), which provide edge blanking and high-speed masking. In contrast, galvo-galvo scanners make it easy to target specific regions within the field of view and offer consistent, user-defined dwell times. By configuring your microscope with both types of scan paths, the user can perform scans optimized for speed or sensitivity with the same instrument. Both scan paths are also required for photostimulation/ uncaging experiments, which use the galvogalvo scan path for photoactivation and the galvo-resonant scan path for high-frame-rate functional imaging. Thorlabsâ&#x20AC;&#x2122; standard galvo-resonant scanner operates at 8 kHz. A 12 kHz version will soon be released that offers higher frame rates. Please contact us at ImagingSales@thorlabs.com for details and availability.

SCANNER

Primary Galvo-Resonant Scan Path Secondary Galvo-Galvo Scan Path Single-Photon Fiber Laser in Secondary Scan Path This Bergamo II microscope has been configured with dual galvo-resonant and galvo-galvo scan paths. Our galvo-galvo scan path (shown in blue) can be augmented with a separate, fibercoupled CW laser (see pages 164 - 167) for photoactivation, preserving power in the primary galvo-resonant scan path (shown in red) for deeper imaging into the specimen.

GALVO-RESONANT

GALVO-GALVO

Scan Speed

30 fps at 512 x 512 Pixels 400 fps at 512 x 32 Pixels

3 fps at 512 x 512 Pixels 48 fps at 512 x 32 Pixels 70 fps at 32 x 32 Pixels

Dwell Time

Varies Across Scan as Function of X (Longer at Scan Edges); Not User-Definable

Identical for All Pixels; User-Definable

Signal to Noise

Higher at Scan Edges

Identical for All Pixels

Scan Geometry

Entire Field of View is Scanned (Can Use Pockels Cell to Mask Regions); Scans Must be Symmetric About Y-Axis

Targeted Illumination

Imaging Systems Bergamo Series Microscopes

Bergamo II Modules in Detail Periscopes Most lasers used in multiphoton microscopy are delivered by a free-space beam. To stay aligned and optimized, the beam path must remain fixed. The Bergamo II’s ability to translate the objective around the focal plane in up to five axes (X, Y, Z, θ, and Elevation) also requires the beam path to translate along the same axes. Bergamo II systems overcome this engineering challenge using articulating periscopes (for rotating systems) or fixed periscopes (for nonrotating systems).

Cerna Series Microscopes

Essentials Kit

G 2X FFV

R+G 2X EFV

Confocal + Microscopes

4X EFV

G G

Rotating Bergamo II systems are outfitted with multi-joint articulating periscopes. This periscope’s design offers the enhanced flexibility needed to allow the entire scanning system to be tilted with respect to the sample.

R +OCT G

R+G

2X FFV 2X FFV

ThorImageLS Software

2X EFV

Systems TDI Digital

2X EFV 4X EFV Microscopy Optical 4X EFV

Tweezers

Non-rotating Bergamo II systems are equipped with fixed periscopes that permit the microscope’s full travel range in X, Y, and Z to be used without compromising the optical performance.

Super Broadband Scan Optics Bergamo II microscopes feature proprietary scan optics that are optimized and corrected for excitation wavelengths within either the 400 - 1100 nm or 600 1400 nm wavelength range, allowing imaging of a wide variety of fluorophores with single- or multiphoton excitation. The choice of wavelength range is made at the time of the order. These broad ranges, extending from the visible well into the near infrared, were chosen to support the latest widely tunable Ti:Sapphire lasers and OPO systems. Because many applications benefit from stimulus light in the visible range, we chose not to sacrifice performance at these critical wavelengths. Our optics take full advantage of the optical designs used in the lowmagnification, high-numerical-aperture objectives that have become the workhorses of multiphoton microscopy. They allow the back aperture of the objective to be filled up to Ø20 mm while providing a scan area that is 1.6X larger than that offered on our original Accera and Bergamo multiphoton microscopes. This expanded area lets you find a region of interest more quickly or simply image more cells at once.

Detectors Bergamo II systems can be configured with up to four PMT detection channels and are fully compatible with industry-standard fluorescence emission filters. Unlike competing designs, Thorlabs’ detector modules make it simple and quick to exchange filters for different measurements, without having to worry about optical alignment. We employ high-sensitivity GaAsP PMTs in our multiphoton systems. These detectors offer high quantum efficiency, aiding in imaging weakly fluorescent or highly photosensitive samples. Two types are available: a cooled PMT, and a noncooled PMT that offers a smaller package size (see pages 158 - 159 for details).

The emission filters and dichroic holders are held behind magnetically sealed doors on the front of the PMT detection module. Removal and exchange takes less than 5 minutes.

Imaging Systems Bergamo Series Microscopes

Bergamo II Modules in Detail

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Multi-Axis Controller with Touchscreen This controller, originally designed for rotating Bergamo II microscope bodies, has been updated to also support non-rotating bodies. It uses knobs to control up to five motorized axes. On rotating systems, a rocker switch changes between fine objective focusing and translation of the elevator base. Each axis can be disabled on an individual basis in order to maintain a location along the desired direction. The integrated touchscreen lets two spatial locations be saved and retrieved locally. An unlimited number of spatial locations can be saved on the computer running ThorImageLS. The touchscreen also reads out the position of every motor.

Objectives Bergamo II microscopes directly accept infinitycorrected objectives with M32 threads. This thread standard is common on the majority of lowmagnification, high-NA objectives used in multiphoton microscopy. (Pages 222 - 225 list popular Nikon and

Olympus objectives stocked by Thorlabs.) Our scan optics completely utilize the optical designs of these specialized objectives, which provide a large field of view and enhanced light-gathering ability. Each Bergamo II microscope is shipped with highly stable, Thorlabs-manufactured mechanical adapters that enable compatibility with RMS- and M25-threaded objectives.

Rigid Stand Sample Holders Thorlabsâ&#x20AC;&#x2122; Rigid Stands, detailed on pages 296 - 300, are rotatable, lockable, low-profile platforms for mounting slides, recording chambers, our Z-axis piezo stages, and custom experimental apparatuses. Each fixture is supported by a solid Ă&#x2DC;1.5" stainless steel post for passive vibrational damping, which is in turn held to the workstation by the red post holder. A locking collar maintains the height of the platform, allowing it to easily rotate into and out of the optical path, and a quick-release mechanism holds the post in place once the desired position is achieved.

MP250-MLSH MP100-RCH1 Recording Chamber Holder

MP150-RCH1 Slide Holder

Insert Holder for Z-Axis Piezo Stages, Breadboards, and Custom Inserts

See Pages 296 - 300 14

Imaging Systems Bergamo Series Microscopes

Bergamo II Modules in Detail

Cerna Series Microscopes

Extended-Field-of-View Collection Optics Deriving more signal from fewer photons is the fundamental goal of any detection system. To this end, we employ ultrasensitive GaAsP PMTs that offer high quantum efficiency. By positioning the PMTs immediately after the objective (a “nondescanned” geometry), light that is scattered by the sample, which therefore appears to originate outside the objective’s field of view, still strikes the PMTs and adds to the collected signal. This is a unique benefit of multiphoton microscopy. Collecting beyond the objective’s design field of view greatly enhances overall detection efficiency when imaging deep in tissue.

Scientific Camera for Widefield Imaging

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Scientific Cameras Our low-noise, scientific-grade CCD cameras (see pages 307 - 322) were designed for full compatibility with Thorlabs’ multiphoton microscopy systems. Useful for widefield and fluorescent microscopy, they are capable of visualizing in vitro and in vivo samples using reflected light and fluorescence emission. They work in conjunction with the epi-fluorescence module to help locate fiducial markers, and they also enable imaging modalities that do not require laser exposure. Thorlabs’ cameras are driven by our internally developed ThorCam software package, and are available in 1.4 megapixel, 4 megapixel, 8 megapixel, and fast-frame-rate versions. Generally speaking, cameras with lower resolution offer higher maximum frame rates. These cameras also feature a separate auxiliary port that permits the image acquisition to be driven by an external electrical trigger signal.

Extended FOV Collection Optics The camera provides the ability to locate fiducial markers using reflected and fluorescent light and permits sample imaging independent from the laser, keeping photodamage to a minimum.

Bergamo II microscopes are also directly compatible with any camera using industrystandard C-mount or CS-mount threads.

1500M-GE 1.4 Megapixel Scientific Camera

Imaging Systems Bergamo Series Microscopes

ThorImageLS Software

Cerna Series Microscopes

ThorImageLS™ is an image acquisition program that offers tight integration with the Bergamo II microscope hardware. Consisting of a series of panels that complement our selection of Bergamo II modules, it is a complete software package for acquiring, visualizing, and analyzing multiphoton images. Its workflow-oriented interface supports single image, Z-stacks, time series, and image streaming acquisitions, and shows the user the measured images in real time.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Because it was developed under the same roof as the Bergamo II microscopes, it provides seamless and intuitive control for the entire microscopy platform. In-line panels are used, for example, to position our motorized XY and XYZ sample holders, adjust the laser power arriving at the sample, set the travel range and step size of a piezo objective holder, and even tune the wavelength of a Coherent Chameleon™ Ti:Sapphire laser. These panels are shown below.

Experimental Techniques n Exponential

Laser Power Ramping with Increasing Sample Depth n Photoactivate a Selected Region of Interest (Galvo-Galvo Systems) n High-Speed Z-Stacks (Piezo Objective Holder)

Equipment Control n Insert/Remove

Dichroic Mirrors to Switch Between Laser Scanning and Widefield Imaging n Average Frames n Change Pixel Dwell Times (Galvo-Galvo Systems) n Tune Laser Wavelength of Coherent’s Chameleon™ Ti:Sapphire Laser

Data Analysis n Reconstruct

3D Z-Stacks n Assign a Color to Each Detection Channel

Main Window (Image Courtesy of Dr. Hajime Hirase and Katsuya Ozawa, RIKEN Brain Science Institute, Wako, Japan)

Imaging Systems Bergamo Series Microscopes

ThorImageLS Software Z-Stack Acquisition Panel The dedicated panel for Z-stack imaging permits the user to define the start of travel, end of travel, and number of steps along the scan. Once entered, this information is used to calculate the Z-stack thickness and volume. A separate fast Z menu panel is enabled when a piezo objective holder is installed. We have designed our own piezo objective holder for the Bergamo II microscopes and also support Physik Instrumenteâ&#x20AC;&#x2122;s (PI) piezo objective holder. Laser Power Control Panel When imaging deeply in vivo, it is often necessary to increase the laser power with the sample depth in order to mantain power at the focal plane. At the same time, it is not necessary to use high laser power closer to the surface to obtain good signal-to-noise. The power control module automatically and exponentially increases the power as the light is focused deeper into the specimen, balancing the competing requirements of high contrast and minimal photodamage.

High-Speed Z-Stack Acquisitions

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

To further prevent photodamage, it is possible to use a standard or high-speed Pockels cell (see page 149 for our selection) with the power control module to perform edge blanking. When using galvo-resonant scanners, the pixel dwell time is longer at the left and right edges of the scan than in the middle, which can lead to overexposure; edge blanking blocks the laser when the scanner is pointing at the edges. It is also possible with our high-speed Pockels cell to define a mask that prevents arbitrary user-specified regions within the field of view from being exposed to the laser.

Laser Power Control

To use this panel, our variable attenuator, our standard Pockels cell, and/or our highspeed Pockels cell beam conditioning modules are required. Before Edge Blanking

After Edge Blanking (Requires Standard or High-Speed Pockels Cell)

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes

Features and Benefits of the Bergamo II Platform +

LASER SCANNING, WIDEFIELD IMAGING, AND TRANSMITTED LIGHT IMAGING

Essentials Kit

- 30 fps at 512 x 512 Pixels R+G - 400 fps at 512 x 32 Pixels +

• User-Defined Line, Square, or Rectangle Scan Geometries

Confocal Microscopes ThorImageLS Software

• High-Frame-Rate Functional Imaging

Galvo-Resonant Only

2X FFV Choice of Single or Dual Scan Paths

OCT Systems

Galvo-Galvo Only

2X FFV

Primary Galvo-Resonant G R and Secondary

TDI Digital Microscopy

2X EFV

2X FFV

2X EFV

• Use Galvo-Galvo Path to Photoactivate a Region of Interest, then Use Galvo-Resonant Path for High-Frame-Rate Functional Imaging

4X EFV

Epi-Illuminator

• Locate Fiducial Markers without Exposure to Laser Light • Choice of Illumination Source - Thorlabs’ High-Power LEDs Provide Bright, Single-Color Illumination - Industry-Standard Liquid Light Guides Illuminate Sample Across the Visible Spectrum

Scientific Camera Port

• Use Reflected or Fluorescent Light to Visualize the Specimen • Compatible with Thorlabs’ Scientific Cameras or Any Camera with Industry-Standard C-Mount or CS-Mount Threads

Piezo Objective Holder Option

• Perform High-Speed Z-Stack Imaging

Motorized Dichroic Mover Option

• Swap Between Laser Scanning and Widefield Imaging Modalities from the Computer

Dodt Contrast and DIC Modules (Non-Rotating Bodies Only)

• User-Installable and Removable Modules that Configure the Microscope for In Vitro or In Vivo - Dodt Contrast + - Laser-Scanned Dodt Contrast - Differential Interference Contrast (DIC) • Motorized Condenser Stage Optimizes Illumination Conditions at the Imaging Plane • Air or High-NA Oil Immersion Condensers from Nikon

OPTICAL PERFORMANCE

Choice of Detectors

2X FFV

R+G

2X EFV 4X EFV • Combines Advantages of Galvo-Resonant and Galvo-Galvo Scanners in One Instrument

R+G

Galvo-Galvo

Optical Tweezers

• Arbitrary User-Defined Line, Square, or Rectangle Scan Geometries 4X EFV • Longer Dwell Times Help Capture Weak Signals Through Integration • 48 fps at 512 x 32 Pixels and 70 fps at 32 x 32 Pixels • Consistent Dwell Times Across Field of View

R+G

2X FFV

R+G

2X FFV

2X EFV

4X EFV

• Full-Field-of-View Collection Optics with 2 PMTs

2X EFV 4X EFV • Extended-Field-of-View Collection Optics with 2 PMTs are Better at Harvesting Signal Photons

Scattered by Thick Tissue • Extended-Field-of-View Collection Optics with 4 PMTs Increase the Number of Fluorescent Tags EFV that can be Detected Simultaneously • Transmitted Light Module with 2 PMTs Collects Signals in Forward Direction, Ideal for Thin or Weakly Fluorescent Samples (Non-Rotating Bodies Only) • Dodt Illumination Modules Accept Forward-Direction Detector (Non-Rotating Bodies Only)

Minimal Distance Between Objective and First Collecting Lens

• Large Collection Angle for Multiphoton Fluorescence Emission • Increased Collection Efficiency

Periscopes

• Maintain Optimal Laser Alignment and Optical Performance over Microscope’s Entire Travel Range - Articulating Periscopes for Rotating Bodies (X, Y, Z, θ, and Elevation) - Fixed Periscopes for Non-Rotating Bodies (X, Y, and Z)

Scan Paths Designed In House

• Super Broadband Correction Range of 400 - 1100 nm or 600 - 1400 nm • Image Fluorophores from the Visible Well into the Infrared with Single- or Multiphoton Excitation • Specifically Designed to Match the Low-Magnification, High-NA Objectives Popularly Used in Multiphoton Microscopy • Take Full Advantage of the Latest Widely Tunable Ti:Sapphire and OPO Systems • Diffraction-Limited Image Quality • Fill Up to a Ø20 mm Back Aperture Objective • Up to F.N. 20

Imaging Systems

Features and Benefits of the Bergamo II Platform

Bergamo Series Microscopes Cerna Series Microscopes

DAY-TO-DAY USAGE Choice of ThorImageLS, ScanImage, or Your Own Software

• Open-Source ThorImageLS: Our Internally Developed Solution • ScanImage: 4.x, 5.x • Full SDK for LabVIEW and C++

Touchscreen Controller

• Touchscreen Shows Current Position of All Axes • Tap to Save and Retrieve Two Positions in Space

Easy-to-Reach Emission Filters and Dichroic Holders

• Filters are Accessed from the Front of the Microscope and Take Less than 5 Minutes to Exchange

Input and Output Triggers

• Use Electrical Signals to Synchronize All Your Equipment • Input Triggers can Start a Single Series or an Indefinite Series • Output Triggers can be Sent at the Beginning of a Frame or a Line • High-Bandwidth Signal Integration with Electrophysiology

Large User-Adjustable Volume Underneath Objective

• Accommodates Large Preps and Setups • Approach Angles Around Objective are Not Restricted • Rotating Bodies have 5" (12.7 cm) of Coarse Z Travel (Elevation) for Easily Adapting the Microscope for Differently Sized Experimental Setups

-5° to +95° or -50° to +50° Rotation of the Focal Plane (Rotating Bodies Only)

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

• Image Different Sections of the Brain Without Having to Move Your Specimen or Refocus the Objective

BEAM CONDITIONING MODULES

R+G • Blank Edges to Prevent Overexposure at the Extremes of the Field of View • High-Speed Option Masks Out Arbitrarily Shaped Regions to Further Limit Exposure

Pockels Cell

2X FFV

2X EFV

4X EFV

Variable Attenuator

• Reduce Laser Power at Sample Using the Computer or Knob • Block Laser Beam Between Measurements

Variable Beam Expander

• Match Objective’s Back Aperture to Utilize the Entire Design FOV • Overfill Objective to Maximize Resolution and Use the Entire NA • Underfill Objective to Increase Power Density and Image Deeper in Thick Preparations

SAMPLE HOLDERS

Rigid Stands for Slides, Recording Chambers, or Platforms

• Minimal Footprint Conserves Space Around the Objective and the Microscope • Slim Profile Leaves Room for Dodt or DIC Imaging Modules (Non-Rotating Bodies Only) • Excellent Long-Term Stability • Easily Rotate Samples Into and Out of the Beam Path

XY Platforms for Micromanipulators

• Large Working Space that Surrounds the Objective on Three Sides • + Ideal for Setups Where the Sample and Apparatus Need to Move in Unison, Such as Patch Clamping • 2" Travel in X and Y; 0.5 µm Encoder Resolution

THORLABS SUPPORT

R+G • Engineers Work Under One Roof to Lower Your Costs and Create Seamless Solutions • Expertise in Every System Component

Fully Designed and Manufactured In House

2X FFV

2X EFV

4X EFV

Modular System Construction

• As Your Experimental Needs Evolve, Upgrade Your Microscope Without Sacrificing Existing Capabilities

Professional Installation

• Thorlabs Technician Visits Your Lab to Assemble, Test, and Demonstrate Use of Your Microscope

Quick Support

• Communicate with Our Support Staff Faster than an Engineer Could Travel to Your Location • Thorlabs Technicians and Application Specialists Available for Videoconferencing • Thorlabs Will Ship You a Camera with a Microphone to Facilitate the Conversation • With Permission, Thorlabs Will Remote Desktop in to Address Software Issues

Imaging Systems Bergamo Series Microscopes

Beam Conditioning Modules

Cerna Series Microscopes

Thorlabs’ Beam Conditioning Modules are add-ons that condition the light emitted by the Ti:Sapphire laser prior to it entering the microscope. These modules provide the following functions:

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

• Pockels Cells provide edge blanking and masking, which help prevent photobleaching and permanent photodamage in samples.

BCM-PA Variable Attenuator

• Variable Beam Expanders are used to match, overfill, or underfill multiphoton objectives, optimizing the experiment for use of the entire field of view, high resolution, or higher transmission into thick samples, respectively. • Variable Attenuators help maintain uniform illumination intensity as the sample depth increases and prevent the sample from seeing the full power of the laser. These modules have dedicated panels in ThorImageLS that provide tight integration with the rest of your microscope hardware. For more details on these modules, please see pages 149 – 151.

BCM-VBE Variable Beam Expander

Beam conditioning modules are inserted into the optical path between the laser source and the microscope. Lens tubes enclose the beam path to serve as protective shielding.

Imaging Systems Bergamo Series Microscopes

Specifications LASER SCANNING Scan Path Wavelength Range

400 - 1100 nm or 600 - 1400 nm

Field of View

20 mm Diagonal Square (Max) at the Intermediate Image Plane One or Two Scan Paths: Galvo-Resonant, Galvo-Galvo, or Both

Scanners

Galvo-Resonant

8 kHz Resonant Scanner (X) and Galvanometric Scan Mirror (Y)

Galvo-Galvo

Two Galvanometric Scan Mirrors (X and Y)

Galvo-Resonant

30 fps at 512 x 512 Pixels 400 fps at 512 x 32 Pixels

Galvo-Galvo

3 fps at 512 x 512 Pixels 48 fps at 512 x 32 Pixels 70 fps at 32 x 32 Pixels Pixel Dwell Time: 0.4 to 20 µs

Scan Speed

Scan Mode

Line, Square, or Rectangle

Scan Zoom

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy

1X to 36X

Scan Resolution

Up to 2048 x 2048 Pixels (Bi-Directional); Up to 4096 x 4096 Pixels (Unidirectional)

Compatible Objective Threadings

M32, M25, and RMS

Optical Tweezers

DETECTION Backward Direction

Two or Four Ultrasensitive GaAsP PMTs

Forward Direction (Non-Rotating Bodies Only)

Two Ultrasensitive Compact GaAsP PMTs Maximum of Four PMTs Controlled by the Software at a Given Time

Collection Optics

Full or Extended Field of View User-Changeable Emission Filters and Dichroic Cubes

Wavelength Range

300 - 720 nm

REFLECTED WIDEFIELD IMAGING Manual or Motorized Switching Between Scanning and Widefield Modes LED or Liquid Light Guide Illumination C-Mount Camera Port for High-Sensitivity CCD TRANSMITTED LIGHT IMAGING (NON-ROTATING BODIES ONLY) User-Installable Dodt Contrast, Laser-Scanned Dodt Contrast, Differential Interference Contrast (DIC), and Motorized Condenser Modules Air or Oil Immersion Condenser TRANSLATION 2" Total Travel; 0.5 µm Encoder Resolution

XY Fine Z

1" Total Travel; 0.1 µm Encoder Resolution

Elevation / Coarse Z (Rotating Bodies Only) Rotation (Rotating Bodies Only)

5" (12.7 cm) Total Travel; 1 µm Encoder Resolution -5° to +95° or -50° to +50° Around Objective Focus 0.1° Encoder Resolution

Additional Options Include XY Platforms, Z-Axis Piezo Stages, and Piezo Objective Holders

Lifetime Support Thorlabs’ team of scientists, engineers, and expert technicians works under one roof, ensuring that the expertise needed to address your pre- and postpurchase concerns is always available. We look forward to helping you determine which system will best meet your imaging needs. To ensure that the intended performance of your Bergamo II system is achieved, a Thorlabs technician will visit your lab to assemble, test, and demonstrate the use of your microscope. After installation, our support staff will be available electronically, faster than an engineer could travel to your location. As part of our included post-installation service package, Thorlabs will include a camera and microphone for videoconferencing, allowing you to show us your experimental setup. With permission, we will also remote desktop in to address software issues. The yellow false-colored image shown on computer screens on pages 3 - 9 is courtesy of Dr. Hajime Hirase and Katsuya Ozawa, RIKEN Brain Science Institute, Wako, Japan.

Bergamo II Series 1500M-GE Thorlabs Scientific Camera WFA4102 0.5X Camera Port

WFA4105 Nikon Y-TV Tube & C-Mount Adapter

WFA4100 1X Camera Port

For EMB3000

High-Power Mounted LEDs

HPLS243 Plasma Light Source X-CITE 200 DC Stabilized Fluorescence Light Source

PSF2000 Only Available for Fixed Periscope Dual Scan Path Systems For EMB1000

Step 16:

Choose an Uncaged Path Fiber Launch

Add a Secondary Path Periscope

Step 14:

Choose Widefield Viewing Accessories

Step 13:

WFA2001 Single Cube Epi Illuminator

Widefield Imaging MCM3000 3-Axis Controller

PLS-X 1D 1" Stage

OPX1100 Primary Imaging Path Galvo/Resonant OPX2200 Secondary Imaging Path Galvo/Galvo (Dual Scan Paths)

Step 12:

Choose Motion Controller(s)

MCM5000 5-Axis Controller (Required for Rotating Bodies)

PLS-XY 2D 1" XY Stage

Motion Control

PMP-2XY(/M) Manipulator XY Platform

OPX4220 PA/Uncaging VIS/NIR

Step 15:

Only Required for Dual Scan Path Systems

MBE74100 Nikon Epi Illuminator (Only Compatible w/ Non-Rotating Bodies and FFV2001 PMT Module)

Widefield Illuminators

ELB4050 Elevator Base w/ XYZ + θ Rotating Body, -50° to +50° (Single Path Only)

For ELB4000

Without Trinoculars WFA4000 Nikon Trinoculars with PSF2100 Eyepieces & IR Filter Fixed Periscope With Trinoculars

This premium configuration provides XYZ + θ motion for the objective, with 5" of coarse Z travel and a rotation range of -5° to +95°. It is equipped with 4 PMTs, extended FOV collection optics, and dual scan paths.

EMB1000 Z-Axis Non-Rotating Body

WFA4101 0.75X Camera Port

WFA4106 Nikon Y-TV Tube & 0.7X C-Mount Adapter

Bergamo II Series Configuration B248

PSA2002 Articulating Periscope

OPX1200 Primary Imaging Path Galvo/Galvo OPX2100 Secondary Imaging Path Galvo/Resonant (Dual Scan Paths) Step 11: Choose a Sample Holder

WFA3000 DIC Turret Adapter Stand & MLS Insert Holder

Nikon MBH76210 DIC Slider 10X Plan Fluor

Stand & Sample Holder

Nikon MBH76220DIC Slider 20X Plan Fluor, Plan APO (N2 Dry)

Stand & Slide Holder

DIC Objective Slider (Choose One Compatible with Your Objective)

This entry-level configuration offers Z-axis motion for the objective, with an XY platform that moves the experimental apparatus. It is equipped with 2 PMTs, full FOV collection optics, and a galvo-galvo scan path.

Nikon MBH76240 DIC Slider 40X Plan Fluor, Plan APO (Air), Plan APO (LWD 40X 1.15 NA) Nikon MBH76225 DIC Slider 25X

Nikon MBH76640 DIC Slider 40X APO NIR (N2)

Nikon MBH76160 DIC Slider 60X APO NIR (N2)

Polarizers & Prisms for DIC Condenser DIC Analyzer

Nikon MBD75300 FN-PT Polarizer Turret for VIS & IR Polarizers

Nikon MBH76720 DIC Slider 16X

Bergamo II Series Configuration B212

Requires MBE74100 from Step 14

DIC Condenser Polarizers Nikon MBN71946 FN-P Visible Rotatable

Nikon MBN71960 FN-IRP IR Rotatable

Nikon MBN71950 FN-IR/FA IR & VIS DIC Analyzer DIC Condenser Prisms

Nikon MBH72300 N2 Dry (10X)

Nikon MBH72310 N2 Dry (16X - 100X)

Multiphoton Microscopes LOM1200 2-Position Manual Objective Mover

LOM2200 2-Position Motorized Objective Mover

WFA5100 Single Objective DIC Prism Holder

DIC Prism Holder (Non-Rotating Bodies Only)

PI-P725 Long-Travel Objective Stage EMB3000 XYZ Non-Rotating Body

PDM1000 Manual Dichroic Mover

FRM2000 750 nm Shortpass

Step 3:

Body Choice Limits Accessory Options

Choose a Primary Reflector/ Dichroic

Step 5:

PMTs with Preamps and PSU

Non-Rotating Bodies Only

Step 6:

Objectives & Holders

Step 7:

OPX1100 Primary Imaging Path Galvo/Resonant (Single Scan Path)

Choose a Reflected PMT Detection Module

Step 8:

OPX1200 Primary Imaging Path Galvo/Galvo (Single Scan Path)

FFV2001 2 Ch Full FOV Module

Non-Rotating Bodies Only

Choose a Condenser

Step 10:

EFV2001 2 Ch Extended FOV Module

Nikon MBL78700 D-CUO DIC Oil 1.4 NA

EFV2050 2 Ch Extended FOV Module w/ Shutter

EFV4001 4 Ch Extended FOV Module

Nikon MBL78600 D-CUD Universal 0.9 NA

WFA0100 Transmitted Light Module Adapter

WFA1100 Dodt/Gradient

WFA1150 Laser Scanned Dodt

Transmitted Light Illumination Kits WFA1020 NIR LED

BSA2000 Compact Condenser Mounting Arm

ZFM2000 Condenser Focus Module *See Controller on Step 12

CSA2000 Condenser Mounting Arm

Non-Rotating Bodies Only TDM1200 2 Ch Full FOV

WFA1010 VIS LED

Nikon MBL70100 FN-C LWD 0.78 NA

Step 9:

Non-Rotating Bodies Only

WFA1000 Transmitted Light

EFV4050 4 Ch Extended FOV Module w/ Shutter

Choose a Transmitted PMT Detection Module

Brightfield Module & Accessories DIC Components (Not Available if Transmitted PMTs are Selected in Step 9)

PMT2100 Compact Non-Cooled GaAsP PMT

PMT2000 Cooled GaAsP PMT

Choose a Dual Objective Nosepiece

Choose Optical Paths

N25X-APO-MP

N40XLWD-NIR N40X-NIR N60X-NIR *Other Objectives Available Upon Request

Step 4:

Step 1:

N20X-PFH

FRM2100 810 nm Shortpass

Choose a Dichroic Mover

Step 2:

Choose a Transmitted Light Module

N16XLWD-PFH

Multiphoton Objectives

Primary Reflectors

Choose a Bergamo II Body

PI-P726 Piezo Scanner for Heavy Objectives

Piezo Stages for Z-Stacks

PDM2000 Motorized Dichroic Mover

FRM1000 Silver-Coated Primary Reflector (Single Path Only) ELB4000 Elevator Base w/ XYZ + θ Rotating Body, -5° to +95°

WFA5110 CFI175 to CFI160 DIC Prism Adapter

PMTs with Preamps & PSU

PMT2100 Compact Non-Cooled GaAsP

Illumination Kit & Detector Included WFA1050 VIS & NIR LED

DC4100 4 Ch 1 Mod Input

LEDD1B 1 Ch

LED Drivers

DC4104 4 Ch 4 Mod Input

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software

Rotating Bergamo II Configurations The flowcharts on the following pages illustrate in detail the modules that comprise a Bergamo II multiphoton microscope and list Thorlabs’ Item Number for each module. They can be used to determine module compatibility and can be combined with the price boxes on pages 32 - 35 to estimate the price of the desired microscope.

Customers interested in a rotating Bergamo II configuration should start on this page. For nonrotating Bergamo II configurations, start on page 27.

Lasers and Related Items Sold Separately Please note that the flowcharts do not include lasers, beam conditioning modules (see pages 149 - 151), or optics for directing the beam into the microscope.

OCT Systems TDI Digital Microscopy

Microscope Body

ELB4050

Optical Tweezers

ELB4000

Elevator Base R +XYZ G+ with θ Body, -50° to +50º

2X FFV

Key

2X EFV

4X EFV

Elevator Base R +XYZ G+ with θ Body, -5° to +95º

2X FFV

2X EFV

4X EFV

Single Scan Path

Independent Choice Dependent Choice

Dual Scan Paths

G Scan Paths

R+G

OPX1100 Primary Galvo-Resonant 2X FFV 2X EFV

OPX1100 G R R+G Primary

OPX1200

R+G

OPX1200 Primary Galvo-Galvo

Galvo-Resonant

OPX2200 FFV 2X EFV 4X2X EFV

Primary

4X EFV Galvo-Galvo 2X FFV 2X EFV

4X EFV OPX2100

Secondary Galvo-Galvo

Secondary Galvo-Resonant

PSA2002

No Periscope

Secondary Scan Path Periscope for XYZ + θ Body

Scan Paths:

FRM1000

Primary Scan Path Reflector / Dichroic

FRM2100 810 nm Shortpass

750 nm Shortpass

MCM5000 Multi-Axis Controller with Touchscreen

Scan Paths:

Collection Optics

PDM2000

Manual Dichroic Mover

FFV2001 2 Channels, Full FOV

MDFM-MF2 Filter Cube

PDM1000

2X FFV Reflected Detection Module:

Fiber-Coupled Input (Laser Not Included)

FRM2000

Silver-Coated Reflector

Motion Controller

Primary Scan Path Dichroic Mover

OPX4220

No Fiber Laser Input

Fiber Laser Input for Secondary Scan Path

R+G G

2X EFV

4X EFV 2X FFV

Motorized Dichroic Mover

R+G 2X EFV

4X EFV 2X FFV

GR + GR

R + GG

R+G

2X 2X EFVFFV4X 2X EFVEFV

4X2XEFV FFV

EFV2001

EFV2050

EFV4001

EFV4050

2 Channels, Extended FOV

2 Channels with Shutter, Extended FOV

4 Channels, Extended FOV

4 Channels with Shutter, Extended FOV

2X EFV

4X EFV

Imaging Systems Bergamo Series Microscopes

Rotating Bergamo II Configurations Reflected Detection Module: PMTs

PMT2000

PMT2100

GaAsP PMT, Cooled with Transimpedance Amplifier

GaAsP PMT, Non-Cooled, Compact with Transimpedance Amplifier

One Needed for Each Channel

Our reflected detection modules are compatible with industry-standard Ø25 mm bandpass filters and 25 mm x 36 mm dichroic mirrors. The selection shown below are some of the most commonly ordered types. Please see pages 228 - 231 for more choices stocked by Thorlabs.

Dichroic Mirrors (One Needed for 2-Channel Systems; Three Needed for 4-Channel Systems)

Bandpass Filters (One Needed for Each Channel) Reflected Detection Module: Bandpass Filters and Dichroic Mirrors

22-0093: 390 nm CWL, 40 nm Bandpass 22-0206: 395 nm CWL, 25 nm Bandpass 22-0055: 447 nm CWL, 60 nm Bandpass 22-0197: 460 nm CWL, 14 nm Bandpass 22-0208: 460 nm CWL, 50 nm Bandpass 22-0210: 525 nm CWL, 40 nm Bandpass 22-0045: 525 nm CWL, 50 nm Bandpass 22-0183: 587 nm CWL, 35 nm Bandpass 22-0066: 593 nm CWL, 46 nm Bandpass 22-0076: 607 nm CWL, 70 nm Bandpass 22-0081: 625 nm CWL, 90 nm Bandpass 22-0184: 630 nm CWL, 92 nm Bandpass 22-0185: 650 nm CWL, 60 nm Bandpass

22-0094: 409 nm Longpass 22-0207: 425 nm Longpass 22-0054: 495 nm Longpass 22-0211: 565 nm Longpass 22-0044: 562 nm Longpass 22-0056: 605 nm Longpass

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

WFA2001 Single-Cube Epi-Illuminator

Epi-Illuminator

MDFM-MF2 Filter Cube The filter sets listed below are some of the most commonly ordered sets. Our full selection is on page 229.

EpiIlluminator:

MDF-BFP: Blue Fluorescent Protein (BFP) Emission, Excitation, and Dichroic Filter Set MDF-GFP2: Alexa Fluor® 488 Emission, Excitation, and Dichroic Filter Set MDF-MCHB: mCherryB Emission, Excitation, and Dichroic Filter Set MDF-TOM: tdTomato Emission, Excitation, and Dichroic Filter Set

Filter Set

High-Power LEDs (Choose 1) EpiIlluminator: Light Source

M470L3

M940L3

470 nm LED

940 nm LED

M565L3

MCWHL5

565 nm LED

Cold White, 6500 K LED

EpiIlluminator:

LEDD1B

Light Source: High-Power LED Driver

TPS001

Widefield Viewing: Camera Tube

Widefield Viewing: Camera

T-Cube LED Driver T-Cube Power Supply

HPLS243 Plasma Light Source with Liquid Light Guide

X-CITE 200 DC Lamp with Liquid Light Guide

DC4100

DC4104

4-Channel LED Driver, 1 Modulation Input

4-Channel LED Driver, 4 Modulation Inputs

DC4100-HUB

Breakout Box

WFA4100

WFA4101

WFA4102

1X Camera Port

0.75X Camera Port

0.5X Camera Port

DCU224M Monochrome CCD Camera

340M-GE

1500M-GE

Fast Frame Rate VGA Scientific Camera

1.4 Megapixel Scientific Camera

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit

Rotating Bergamo II Configurations Objectives: Piezo Objective Holder for Fast Z-Stacks

No Piezo Objective Holder

PI-P725

PI-P726

Long-Travel Objective Stage, 400 µm Travel Range

Piezo Scanner for Heavy Objectives, 100 µm Travel Range

Confocal Microscopes ThorImageLS Software

SC-IMG2 Scan NI Package

Bergamo II microscopes directly accept M32-threaded objectives. Each microscope also includes adapters that enable compatibility with RMS- and M25-threaded objectives. The objectives shown below are some available from Thorlabs; please contact us to request others.

OCT Systems TDI Digital Microscopy

SC-IMG2 Scan NI Package

Objectives

Optical Tweezers

N16XLWD-PF: Nikon CFI LWD Plan Fluorite Objective, 16X N20X-PFH: Olympus XLUMPLFLN Objective, 20X N25X-APO-MP: Nikon CFI APO Objective, 25X N40X-NIR: Nikon CFI APO Objective, 40X N40XLWD-NIR: Nikon CFI APO LWD Objective, 40X N60X-NIR: Nikon CFI APO Objective, 60X LUMFLN60XW: Olympus LUMPFLN Objective, 60X

Rigid Stands with Platforms MP100: Rigid Stand with M6-Tapped Platform, 148.1 - 208.5 mm Tall MP150: Rigid Stand with M6-Tapped Platform, 198.1 - 309.3 mm Tall MP200: Rigid Stand with M6-Tapped Platform, 248.1 - 410.1 mm Tall MP250: Rigid Stand with M6-Tapped Platform, 265.9 - 510.9 mm Tall (1/4"-20 Versions Available Upon Request) Rigid Stands with Slide Holders

Sample Holders: Rigid Stands (See Pages 296 - 300 for Full Presentation)

MP100-RCH2: Rigid Stand with Slide Holder, 148.1 - 208.5 mm Tall MP150-RCH2: Rigid Stand with Slide Holder, 198.1 - 309.3 mm Tall MP200-RCH2: Rigid Stand with Slide Holder, 248.1 - 410.1 mm Tall MP250-RCH2: Rigid Stand with Slide Holder, 265.9 - 510.9 mm Tall

Rigid Stands with Recording Chamber Holders

MP100-RCH1: Rigid Stand with Recording Chamber Holder, 148.1 - 208.5 mm Tall MP150-RCH1: Rigid Stand with Recording Chamber Holder, 198.1 - 309.3 mm Tall MP200-RCH1: Rigid Stand with Recording Chamber Holder, 248.1 - 410.1 mm Tall MP250-RCH1: Rigid Stand with Recording Chamber Holder, 265.9 - 510.9 mm Tall

Rigid Stands with Insert Holders

MP100-MLSH : Rigid Stand with Insert Holder, 148.1 - 208.5 mm Tall MP150-MLSH: Rigid Stand with Insert Holder, 198.1 - 309.3 mm Tall MP200-MLSH: Rigid Stand with Insert Holder, 248.1 - 410.1 mm Tall MP250-MLSH: Rigid Stand with Insert Holder, 265.9 - 510.9 mm Tall

PLS-X Sample Holders: Motion Control Options

PLS-XY

One-Axis Motorized Stage with 1" Travel in X

Two-Axis Motorized Stage with 1" Travel in X and Y

MCM3000 3-Axis Controller

Imaging Systems Bergamo Series Microscopes

Non-Rotating Bergamo II Configurations

Cerna Series Microscopes

Microscope Body

EMB3000

EMB1000

(See Page 29 for XY Platform)

Z Body

2X FFV Motion Controller

Essentials Kit

2X EFV

2X FFV

4X EFV

Confocal Microscopes

XYZ Body

R+G

2X EFV

4X EFV

ThorImageLS Software OCT Systems

MCM5000

MCM3000

Multi-Axis Controller with Touchscreen

3-Axis Controller

Single Scan Path

TDI Digital Microscopy Optical Tweezers

Dual Scan Paths Key

R+G

OPX1100 Primary Galvo-Resonant 2X FFV 2X EFV

OPX1100 G R

R+G

Primary Galvo-Resonant

OPX1200

R+G

OPX2200 FFV 2X EFV 4X2X EFV

Primary

4X EFV Galvo-Galvo 2X FFV 2X EFV

Primary Galvo-Galvo Secondary Galvo-Resonant

For XYZ Body Only

For Z Body Only No Periscope

PSF2100

PSF2000

Secondary Scan Path Periscope for XYZ Body

Secondary Scan Path Periscope for Z Body

Scan Paths: Fiber Laser Input for Secondary Scan Path Scan Paths: Primary Scan Path Reflector / Dichroic Scan Paths: Primary Scan Path Dichroic Mover

OPX4220

No Fiber Laser Input

Fiber-Coupled Input (Laser Not Included)

FRM2100

FRM1000

810 nm Shortpass

FRM2000

Silver-Coated Reflector

750 nm Shortpass + +

PDM1000

GManualRDichroicRMover +G G 2X FFV

FFV2001 Reflected Detection Module

Dependent Choice

OPX1200

4X EFV OPX2100

Secondary Galvo-Galvo

Scan Paths

Independent Choice

2 Channels, Full FOV

MDFM-MF2 Filter Cube

2X EFV

4X EFV 2X FFV

PDM2000

R+G 2X EFV

4X EFV 2X FFV

GR + GR

R + GG

Motorized Dichroic Mover

R+G

2X 2X EFVFFV4X 2X EFVEFV

4X2XEFV FFV

EFV2001

EFV2050

EFV4001

EFV4050

2 Channels, Extended FOV

2 Channels with Shutter, Extended FOV

4 Channels, Extended FOV

4 Channels with Shutter, Extended FOV

2X EFV

4X EFV

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit

Non-Rotating Bergamo II Configurations PMT2100

GaAsP PMT, Cooled

One Needed for Each Channel

GaAsP PMT, Non-Cooled, Compact

PMTs

Confocal Microscopes ThorImageLS Software

PMT2000

Reflected Detection Module:

Dichroic Mirrors (One Needed for 2-Channel Systems; Three Needed for 4-Channel Systems)

Bandpass Filters (One Needed for Each Channel)

OCT Systems

Reflected Detection Module:

TDI Digital Microscopy

Bandpass Filters and Dichroic Mirrors

Optical Tweezers

22-0094: 409 nm Longpass 22-0207: 425 nm Longpass 22-0054: 495 nm Longpass 22-0211: 565 nm Longpass 22-0044: 562 nm Longpass 22-0056: 605 nm Longpass

Compatible with All Reflected PMT Modules Epi-Illuminator

Required for DIC; Only Compatible with FFV2001 MBE74100

WFA2001 Single-Cube Epi-Illuminator

Nikon Epi-Illuminator for 6 Dichroics

MDFM-MF2

MDFM-TE2000 Filter Cube (Sold Individually)

Filter Cube

The filter sets listed below are some of the most commonly ordered sets. Our full selection is on page 229.

EpiIlluminator: Filter Set

EpiIlluminator: Light Source

EpiIlluminator: Light Source: High-Power LED Driver

Widefield Viewing: Trinoculars

High-Power LEDs (Choose 1) M470L3 M940L3 470 nm LED

940 nm LED

M565L3

MCWHL5

565 nm LED

Cold White, 6500 K LED

HPLS243

DC4100

LEDD1B T-Cube LED Driver

TPS001 T-Cube Power Supply

WFA4000 Nikon Trinoculars

X-CITE 200

Plasma Light Source with Liquid Light Guide

DC Lamp with Liquid Light Guide

DC4104

4-Channel LED Driver, 1 Modulation Input

4-Channel LED Driver, 4 Modulation Inputs

DC4100-HUB

Breakout Box

No Trinoculars

Imaging Systems Bergamo Series Microscopes

Non-Rotating Bergamo II Configurations Widefield Viewing: Camera Tube

No Camera Tube

WFA4105

WFA4106

Nikon Y-TV Tube with C-Mount

Nikon Y-TV Tube with 0.7X C-Mount

WFA4100

WFA4101

WFA4102

1X Camera Port

0.75X Camera Port

0.5X Camera Port

Cerna Series Microscopes Essentials Kit Confocal Microscopes

Widefield Viewing: Cameras

No Camera

DCU224M

340M-GE

1500M-GE

Monochrome CCD Camera

Fast Frame Rate VGA Scientific Camera

1.4 Megapixel Scientific Camera

ThorImageLS Software OCT Systems TDI Digital Microscopy

Objectives: Piezo Objective Holder

PI-P725 None

PI-P726

400 µm Long-Travel Objective Stage

100 µm Travel Range Piezo Scanner for Heavy Objectives

SC-IMG2

Optical Tweezers

SC-IMG2

Scan NI Package

Scan NI Package Bergamo II microscopes directly accept M32-threaded objectives. Each microscope also includes adapters that enable compatibility with RMS- and M25-threaded objectives. The objectives shown below are some available from Thorlabs; please contact us to request others.

Objectives

Objectives: Dual Objective Nosepieces (Optional)

Condensers: Mounting Arms, Focusing Module, and Controller

N16XLWD-PF: Nikon CFI LWD Plan Fluorite Objective, 16X N20X-PFH: Olympus XLUMPLFLN Objective, 20X N25X-APO-MP: Nikon CFI APO Objective, 25X N40XNIR: Nikon CFI APO Objective, 40X N40XLWD-NIR: Nikon CFI APO LWD Objective, 40X N60X-NIR: Nikon CFI APO Objective, 60X LUMFLN60XW: Olympus LUMPFLN Objective, 60X

Requires PDM2000

LOM1200 Manual 2-Position Mover

LOM2200 Motorized 2-Position Mover

MBL70100

MBL78600

Nikon FN-C LWD Condenser, 0.78 NA

Nikon D-CUD Condenser, 0.9 NA

MBL78700 Nikon D-CUO DIC Oil Condenser, 1.4 NA

BSA2000

CSA2000

Compact Condenser Mounting Arm with Coarse Adjustment

Condenser Mounting Arm with ±2 mm Travel in X and Y

ZFM2000 Focusing Module with 1" Motorized Travel

MCM3000 3-Axis Controller

Transmitted Detection Module Transmitted Detection Module: PMTs

TDM1200 2 Channels, Full FOV

One Needed for Each Channel

PMT2100 GaAsP PMT, Non-Cooled, Compact

Imaging Systems Bergamo Series Microscopes

Non-Rotating Bergamo II Configurations

Cerna Series Microscopes

WFA0100

Essentials Kit Confocal Microscopes

Transmitted Light Module Adapter

WFA1000 Transmitted Light Illumination

ThorImageLS Software

Transmitted Light and DIC Imaging Module

Transmitted Light Only

Optical Tweezers

Transmitted Light Illumination: Illumination Kits

WFA1010

WFA1020

Visible LED Kit

Laser-Scanned Dodt Contrast Module

Dodt Contrast Module

DIC Imaging (Requires MBE74100)

OCT Systems TDI Digital Microscopy

WFA1150

WFA1100

NIR LED Kit

WFA1050 Visible and NIR LED Kit

LEDD1B Transmitted Light Illumination:

Illumination Kit Driver

1-Channel T-Cube LED Driver

DC4100

TPS001

4-Channel LED Driver, 1 Modulation Input

T-Cube Power Supply

DC4100-HUB

Two of Each Required for WFA1050 Only

Breakout Box

DC4104 4-Channel LED Driver, 4 Modulation Inputs

DC4100-HUB Breakout Box

WFA5100

WFA5110

Single-Objective Prism Holder

CFI175 to CFI160 Prism Adapter

WFA3000 Turret Adapter

MBN71950 Visible and NIR DIC Analyzer

Transmitted Light Illumination: DIC Components

MBD75300 Polarizer Turret

MBN71946

MBN71960

Visible Polarizer

NIR Polarizer

MBH72300 Dry DIC Condenser Prism, 10X

MBH72310 Dry DIC Condenser Prism, 16X - 100X

NikonNikon MBH76225 NikonNikon MBH76210 NikonMBH76220DIC MBH76720 Nikon MBH76220DIC Nikon MBH76225 Nikon MBH76225 Nikon MBH76210 Nikon MBH76210 Nikon MBH76720 Nikon MBH76720 Nikon MBH76225 Nikon MBH76225 Nikon MBH76225 Nikon MBH76210 Nikon MBH76210 Nikon MBH76210 Nikon MBH76220DIC Nikon MBH76220DIC Nikon MBH76720 Nikon MBH76720 Nikon MBH76720 Nikon MBH76220DIC Nikon Nikon MBH76220DIC MBH76225 MBH76210 Nikon MBH76720 Nikon MBH76220DIC MBH76210 MBH76720 MBH76220 MBH76225 DIC25X Slider DIC 25XSlider 25X DIC10X SliderDIC 10XSlider DIC 16X 20XSlider Plan DIC Slider 25X Slider 10X DIC Slider16X 10XSliderSlider DIC 16X DICSlider Slider DIC 25XSlider 25X DIC Slider DIC Slider DIC 10XSliderDIC 10X DIC Slider 20X Slider Plan 20X Plan DIC Slider DIC 16XSlider 16X DIC10X Slider 16X Slider 20X Slider Plan 20XSlider Plan Slider DIC Slider 25X DIC Slider DIC 16X20X Plan Slider Plan DICPlan Slider, 10X, DIC Slider, 16X DIC Slider, 20X, for20X Plan Plan Fluor DIC Slider, 25X Fluor, APO (N Fluor PlanFluor, Fluor Fluor Planfor FluorPlanPlan Fluor Fluor, Plan APOAPO (NPlan Dry) Fluor, Plan APO (N Dry) Fluor, Plan APO (N Plan APO Fluor, (N Dry) Dry) 2 Dry) Plan Fluor Fluor, APO (N 2 (N 2 Dry) 2 Plan 2Plan 2 Dry)2 Plan Fluorite Fluorite or Plan Apo (N2 Dry)

Choose One NikonNikon MBH76160 Nikon MBH76640 Nikon MBH76240 Nikon MBH76160 Nikon MBH76160 Nikon MBH76160 Nikon MBH76160 Nikon MBH76160 Nikon MBH76640 Nikon MBH76640 Nikon MBH76240 Nikon MBH76240 Nikon MBH76640 Nikon MBH76640 Nikon MBH76640 Nikon MBH76240 Nikon MBH76240 Nikon MBH76240 MBH76160 Nikon MBH76640 Nikon MBH76240 MBH76240 MBH76640 MBH76160 Compatible with DIC60X Slider 60X DIC Slider 40X DIC40X Slider 40X Plan Fluor, DIC Slider 60X DIC Slider DIC Slider DIC 60X Slider 60X DIC Slider DIC Slider 40X DIC Slider 40X DIC Slider 40X Plan Fluor, DIC Slider 40X Plan Fluor, DIC Slider DIC 40X Slider 40X DIC Slider 40X DIC Slider DIC 40XSlider Plan Fluor, 40X DIC Plan Slider Fluor, Plan Fluor, DIC Slider 60X 60X DIC Slider 40X DIC Slider 40X Plan Fluor, DIC Slider, 40X, for Plan DIC Slider, 40X, for Plan DIC Slider, APO NIR (N ) YourPlan Objective APO NIR Plan APO (Air), Plan APO NIR (N22))(N2)APO NIR APO NIR (N2) 60X, for Plan APO NIR NIR (N (N NIR APO NIR (N(N APO (N )(N 2 (N PlanPlan APO (Air), Plan APO Plan APO(N (Air), Plan APO APO NIR (N APO NIR NIR APO (Air), Plan Plan APO APO (Air), Plan APO APO (Air), Plan 22)) APO NIR (NNIR 2) 2)APO Plan APO (Air), Plan APO 2APO 2)APO 2) APO 2) 2) (LWD 40X 1.15 NA) Fluorite or Plan Apo (Air) 40X40X 1.15 NA) 40X NA) 1.15Apo NA) NIR (N2 Dry) (LWD 40X 1.15 (LWDNA) 40X (LWD 1.15 (LWD NA) 1.15 NA)(LWD (LWD 40X 1.15

Apo NIR (N2 Dry)

Imaging Systems Bergamo Series Microscopes

Non-Rotating Bergamo II Configurations

Cerna Series Microscopes

Rigid Stands with Platforms

MP100: Rigid Stand with M6-Tapped Platform, 148.1 - 208.5 mm Tall MP150: Rigid Stand with M6-Tapped Platform, 198.1 - 309.3 mm Tall MP200: Rigid Stand with M6-Tapped Platform, 248.1 - 410.1 mm Tall MP250: Rigid Stand with M6-Tapped Platform, 265.9 - 510.9 mm Tall (1/4"-20 Versions Available Upon Request)

Essentials Kit Confocal Microscopes ThorImageLS Software

Rigid Stands with Slide Holders

OCT Systems MP100-RCH2: Rigid Stand with Slide Holder, 148.1 - 208.5 mm Tall MP150-RCH2: Rigid Stand with Slide Holder, 198.1 - 309.3 mm Tall MP200-RCH2: Rigid Stand with Slide Holder, 248.1 - 410.1 mm Tall MP250-RCH2: Rigid Stand with Slide Holder, 265.9 - 510.9 mm Tall

Sample Holders: Rigid Stands (See Pages 296 - 300 for Full Presentation)

Rigid Stands with Recording Chamber Holders

TDI Digital Microscopy Optical Tweezers

Rigid Stands with Insert Holders

MP100-MLSH: Rigid Stand with Insert Holder, 148.1 - 208.5 mm Tall MP150-MLSH: Rigid Stand with Insert Holder, 198.1 - 309.3 mm Tall MP200-MLSH: Rigid Stand with Insert Holder, 248.1 - 410.1 mm Tall MP250-MLSH: Rigid Stand with Insert Holder, 265.9 - 510.9 mm Tall

PMP-2XY(/M) XY Platform with 2" Travel in X and Y and 1/4"-20 (M6) Taps

Sample Holders:

PLS-X

PLS-XY

Motion Control Options

One-Axis Motorized Stage with 1" Travel in X

Two-Axis Motorized Stage with 1" Travel in X and Y

Choose One Controller

Motion Controllers

MCM3000 3-Axis Controller

MCM5000 Multi-Axis Controller with Touchscreen

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Bergamo II Pricing The boxes on the following pages contain the prices of the components of a Bergamo II microscope. They should be used in conjunction with the flowcharts on pages 24 - 31 to determine possible configuration options. Do not, for example, attempt to simply pick one item from each box; doing so is likely to result in an incomplete or impossible configuration. If you would like to walk through the options with a representative, we are available at ImagingSales@thorlabs.com. Microscope Body (Step 2 on Pages 22 - 23) ITEM # EMB1000

PRICE 6,900.00

DESCRIPTION Z Body

EMB3000

17,250.00

XYZ Body

ELB4050

43,700.00

XYZ + θ Body, -50° to +50° (Requires MCM5000)

ELB4000

43,700.00

XYZ + θ Body, -5° to +95° (Requires MCM5000)

Motion Controllers (Step 12 on Pages 22 - 23) ITEM # MCM3000

PRICE 4,500.00

MCM5000

10,350.00

DESCRIPTION 3-Axis Controller Multi-Axis Controller with Touchscreen

Scan Paths (Step 1 on Pages 22 - 23) ITEM # OPX1100

PRICE 86,250.00

DESCRIPTION Primary Galvo-Resonant Scan Path

OPX1200

83,950.00

Primary Galvo-Galvo Scan Path

OPX2100

63,250.00

Secondary Galvo-Resonant Scan Path (Requires OPX1200 Primary Galvo-Galvo Scan Path)

OPX2200

60,950.00

Secondary Galvo-Galvo Scan Path (Requires OPX1100 Primary Galvo-Resonant Scan Path)

PSF2000

814.84

PSF2100

1,594.38

Secondary Scan Path Periscope for XYZ Body (Item # EMB3000)

PSA2002

10,500.00

Secondary Scan Path Periscope for XYZ + θ Body, -5° to +95° (Item # ELB4000)

Secondary Scan Path Periscope for Z Body (Item # EMB1000)

Scan Paths: Primary Scan Path Reflector / Dichroic (Step 3 on Pages 22 - 23) ITEM # FRM1000

PRICE 750.00

DESCRIPTION Silver-Coated Reflector

FRM2000

1,450.00

750 nm Shortpass Dichroic Mirror

FRM2100

1,450.00

810 nm Shortpass Dichroic Mirror

Scan Paths: Primary Scan Path Dichroic Mover (Step 4 on Pages 22 - 23) ITEM # PDM1000

PRICE 2,500.00

PDM2000

5,750.00

DESCRIPTION Manual Dichroic Mover Motorized Dichroic Mover

Scan Paths: Fiber Laser Input for Secondary Scan ITEM # OPX4220

PRICE 5,100.00

Path*

(Step 16 on Pages 22 - 23)

DESCRIPTION Single-Photon Fiber Laser Input for Photoactivation/Uncaging

*Laser Not Included

Objectives (Step 6 on Pages 22 - 23) ITEM # N16XLWD-PF

PRICE 5,533.00

DESCRIPTION Nikon CFI LWD Plan Fluorite Objective, 16X

N20X-PFH

6,706.00

Olympus XLUMPLFLN Objective, 20X

N25X-APO-MP

22,336.00

Nikon CFI APO Objective, 25X

N40X-NIR

2,318.00

Nikon CFI APO Objective, 40X

N40XLWD-NIR

12,973.00

Nikon CFI APO LWD Objective, 40X

N60X-NIR

3,694.00

Nikon CFI APO Objective, 60X

LUMFLN60XW

9,085.00

Olympus LUMPFLN Objective, 60X

Objectives: Piezo Objective Holders for Fast Z-Stacks (Step 6 on Pages 22 - 23) ITEM # PI-P725

PRICE 15,499.00

DESCRIPTION Physik Instrumente (PI) Long-Travel Objective Stage, 400 µm Travel Stage (Requires SC-IMG2)

PI-P726

15,090.00

Physik Instrumente (PI) Piezo Scanner for Heavy Objectives, 100 µm Travel Stage (Requires SC-IMG2)

SC-IMG2

1,500.00

National Instruments Card Package

Objectives: Dual Objective Nosepieces* (Step 5 on Pages 22 - 23) ITEM # LOM1200

PRICE 3,681.11

LOM2200

5,313.26

*Requires PDM2000 Motorized Objective Mover

DESCRIPTION Manual 2-Position Objective Mover Motorized 2-Position Objective Mover (Requires 1 Available Port on MCM3000)

Imaging Systems Bergamo Series Microscopes

Bergamo II Pricing Reflected Detection Modules: Collection Optics (Step 7 on Pages 22 - 23) ITEM # FFV2001

PRICE 4,225.00

DESCRIPTION 2-Channel, Full FOV Detector Module (Requires MDFM-MF2)

MDFM-MF2

473.89

EFV2001

12,500.00

2-Channel, Extended FOV Detector Module

EFV2050

16,000.00

2-Channel, Extended FOV Detector Module with Mechanical Shutter

EFV4001

22,200.00

4-Channel, Extended FOV Detector Module

EFV4050

29,200.00

4-Channel, Extended FOV Detector Module with Mechanical Shutter

Filter Cube for 2-Channel, Full FOV Detector Module

Reflected Detection Modules: PMTs* (Step 7 on Pages 22 - 23) ITEM # PMT2000

PRICE 6,750.00

PMT2100

6,200.00

DESCRIPTION GaAsP PMT with Transimpedance Amplifier, Cooled GaAsP PMT with Transimpedance Amplifier, Non-Cooled Compact

*One Needed for Each Channel

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy

Reflected Detection Modules: Bandpass Filters* ITEM # 22-0093

PRICE 299.00

DESCRIPTION 390 nm CWL, 40 nm Bandpass

22-0206

325.00

395 nm CWL, 25 nm Bandpass

22-0055

299.00

447 nm CWL, 60 nm Bandpass

22-0197

299.00

460 nm CWL, 14 nm Bandpass

22-0208

475.00

460 nm CWL, 50 nm Bandpass

22-0210

349.00

525 nm CWL, 40 nm Bandpass

22-0045

299.00

525 nm CWL, 50 nm Bandpass

22-0183

349.00

587 nm CWL, 35 nm Bandpass

22-0066

299.00

593 nm CWL, 46 nm Bandpass

22-0076

299.00

607 nm CWL, 70 nm Bandpass

22-0081

899.00

625 nm CWL, 90 nm Bandpass

22-0184

299.00

630 nm CWL, 92 nm Bandpass

22-0185

349.00

650 nm CWL, 60 nm Bandpass

Optical Tweezers

*One Needed for Each Channel

Reflected Detection Modules: Dichroic Mirrors* ITEM # 22-0094

PRICE 249.00

DESCRIPTION 409 nm Longpass Mirror

22-0207

225.00

425 nm Longpass Mirror

22-0054

249.00

495 nm Longpass Mirror

22-0211

225.00

565 nm Longpass Mirror

22-0044

249.00

562 nm Longpass Mirror

22-0056

249.00

605 nm Longpass Mirror

*One Needed for 2-Channel Systems; Three Needed for 4-Channel Systems

Sample Holders: Rigid Stands* (Step 11 on Pages 22 - 23) ITEM # MP100-RCH2

PRICE 752.91

DESCRIPTION Rigid Stand with Slide Holder, Adjustment Height: 148.1 - 208.5 mm

MP150-RCH2

766.24

Rigid Stand with Slide Holder, Adjustment Height: 198.1 - 309.3 mm

MP200-RCH2

779.58

Rigid Stand with Slide Holder, Adjustment Height: 248.1 - 410.1 mm

MP250-RCH2

792.91

Rigid Stand with Slide Holder, Adjustment Height: 265.9 - 510.9 mm

MP100-RCH1

648.22

Rigid Stand with Recording Chamber Holder, Adjustment Height: 148.1 - 208.5 mm

MP150-RCH1

661.56

Rigid Stand with Recording Chamber Holder, Adjustment Height: 198.1 - 309.3 mm

MP200-RCH1

674.89

Rigid Stand with Recording Chamber Holder, Adjustment Height: 248.1 - 410.1 mm

MP250-RCH1

688.22

Rigid Stand with Recording Chamber Holder, Adjustment Height: 265.9 - 510.9 mm

MP100-MLSH

769.84

Rigid Stand with Insert Holder, Adjustment Height: 148.1 - 208.5 mm

MP150-MLSH

783.18

Rigid Stand with Insert Holder, Adjustment Height: 198.1 - 309.3 mm

MP200-MLSH

796.51

Rigid Stand with Insert Holder, Adjustment Height: 248.1 - 410.1 mm

MP250-MLSH

809.84

Rigid Stand with Insert Holder, Adjustment Height: 265.9 - 510.9 mm

*See Pages 296 - 300 for Full Presentation

Sample Holders: Motion Control Options (Step 11 on Pages 22 - 23) ITEM # PLS-X PLS-XY PMP-2XY

METRIC ITEM # –

PRICE 1,296.80

DESCRIPTION 1D Stage with 1" Travel in X for Rigid Stands (Requires MCM3000)

–

2,370.52

2D Stage with 1" Travel in X and Y for Rigid Stands (Requires MCM3000)

PMP-2XY/M

12,211.72

XY Platform with 2" Travel in X and Y and 1/4"-20 (M6) Taps (Requires Motion Controller)

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes

Bergamo II Pricing Epi-Illuminator (Step 13 on Pages 22 - 23) ITEM # WFA2001

PRICE 2,300.00

Essentials Kit

MDFM-MF2

473.89

Confocal Microscopes

MDFM-TE2000

ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

MBE74100 $ 2,797.00 $

DESCRIPTION Single-Cube Epi-Illuminator (Requires MDFM-MF2) Filter Cube for Single-Cube Epi-Illuminator Nikon Epi-Illuminator for 6 Filter Cubes (Requires at Least One MDFM-TE2000, Only Compatible with FFV2001 Reflected Detection Module)

466.60

Filter Cube for Nikon Epi-Illuminator

Epi-Illuminator: Filter Set ITEM # MDF-BFP

PRICE 625.00

DESCRIPTION Blue Fluorescent Protein (BFP) Emission, Excitation, and Dichroic Filter Set

MDF-GFP2

745.00

Alexa FluorÂ® 488 Emission, Excitation, and Dichroic Filter Set

MDF-MCHB

745.00

mCherryB Emission, Excitation, and Dichroic Filter Set

MDF-TOM

745.00

tdTomato Emission, Excitation, and Dichroic Filter Set

Epi-Illuminator: Light Source: High-Power LEDs* (Step 13 on Pages 22 - 23) ITEM # M470L3

PRICE 260.00

DESCRIPTION 470 nm LED

M565L3

205.00

M940L3

205.00

940 nm LED

MCWHL5

187.51

Cold White, 6500 K LED

565 nm LED

*Requires Driver

Epi-Illuminator: Light Source: High-Power LED Driver (Step 13 on Pages 22 - 23) ITEM # LEDD1B

PRICE 284.00

DESCRIPTION 1-Channel T-Cube LED Driver (Requires TPS001)

TPS001

25.00

DC4100

2,505.00

4-Channel LED Driver, 1 Modulation Input (Requires DC4100-HUB)

DC4104

2,759.00

4-Channel LED Driver, 4 Modulation Inputs (Requires DC4100-HUB)

DC4100-HUB

300.00

Power Supply for 1-Channel T-Cube LED Driver

Breakout Box for 4-Channel LED Drivers

Epi-Illuminator: Light Source: Liquid Light Guide (Step 13 on Pages 22 - 23) ITEM # HPLS243

PRICE 3,810.00

X-CITE 200

6,495.00

DESCRIPTION Plasma Light Source with Liquid Light Guide DC Lamp with Liquid Light Guide

Widefield Viewing: Trinoculars (Step 14 on Pages 22 - 23) ITEM # WFA4000

PRICE 2,850.00

DESCRIPTION Nikon Trinoculars with Eyepieces

Widefield Viewing: Camera Tube (Step 14 on Pages 22 - 23) ITEM # WFA4100

PRICE 1,450.00

DESCRIPTION 1X Camera Tube for WFA2001 Single-Cube Epi-Illuminator

WFA4101

1,450.00

WFA4102

1,450.00

WFA4105

387.00

Nikon Y-TV Tube with C-Mount for MBE74100 Nikon Epi-Illuminator

WFA4106

764.00

Nikon Y-TV Tube with 0.7X C-Mount for MBE74100 Nikon Epi-Illuminator

0.75X Camera Tube for WFA2001 Single-Cube Epi-Illuminator 0.5X Camera Tube for WFA2001 Single-Cube Epi-Illuminator

Widefield Viewing: Cameras (Step 14 on Pages 22 - 23) ITEM # DCU224M

PRICE 2,184.00

DESCRIPTION Monochrome CCD Camera

340M-GE

4,400.00

Fast-Frame-Rate VGA Scientific Camera

1500M-GE

5,100.00

1.4 Megapixel Scientific Camera

Condensers (Step 8 on Pages 22 - 23)

ITEM # MBL70100

PRICE 1,943.00

DESCRIPTION Nikon FN-C LWD Condenser, 0.78 NA

MBL78600

1,836.00

Nikon D-CUD Condenser, 0.9 NA

MBL78700

2,227.00

Nikon D-CUO DIC Oil Condenser, 1.4 NA

Imaging Systems Bergamo Series Microscopes

Bergamo II Pricing Condensers: Condenser Mounting Arm (Step 8 on Pages 22 - 23) ITEM # BSA2000

PRICE 850.00

CSA2000

678.00

DESCRIPTION Compact Condenser Mounting Arm with Coarse Adjustment Condenser Mounting Arm with Â±2 mm Travel in X and Y

Condensers: Condenser Focusing Module and Controller (Step 8 on Pages 22 - 23) ITEM # ZFM2000

PRICE 2,050.00

MCM3000

4,500.00

DESCRIPTION Focusing Module with 1" Motorized Travel (Requires 1 Available Port on MCM3000) 3-Axis Controller

Transmitted Detection Modules (Step 9 on Pages 22 - 23) ITEM #

PRICE

DESCRIPTION 2-Channel, Full FOV Detector Module, Forward Direction (Requires MDFM-MF2, PMTs Sold Separately, Not Compatible with PMT2000)

TDM1200 $ 15,000.00 MDFM-MF2

473.89

PMT2100

6,200.00

Filter Cube for 2-Channel, Full FOV Detector Module

WFA1000 WFA1100 WFA1150

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy

GaAsP PMT, Non-Cooled, Compact (One Needed for Each Channel) Optical Tweezers

Transmitted Light Illumination (Step 10 on Pages 22 - 23) ITEM # WFA0100

Cerna Series Microscopes

PRICE 523.16

DESCRIPTION Transmitted Light Module Adapter (Required for All Following Modules)

4785.78

Transmitted Light and DIC Imaging Module (Requires Illumination Kit)

5,008.00

Dodt Contrast Module (Requires Illumination Kit)

6,819.82

Laser-Scanned Dodt Contrast Module (Includes Illumination Kit and Detector)

Transmitted Light Illumination: Illumination Kits* (Step 10 on Pages 22 - 23) ITEM # WFA1010

PRICE 750.83

WFA1020

768.83

WFA1050

1,308.13

DESCRIPTION Visible LED Kit NIR LED Kit Visible and NIR LED Kit

*Requires Driver

Transmitted Light Illumination: Illumination Kit Driver (Step 10 on Pages 22 - 23) ITEM # LEDD1B

PRICE 284.00

TPS001

25.00

DC4100

2,505.00

4-Channel LED Driver, 1 Modulation Input (Requires DC4100-HUB)

DC4104

2,759.00

4-Channel LED Driver, 4 Modulation Inputs (Requires DC4100-HUB)

DC4100-HUB

300.00

Transmitted Light Illumination: DIC

DESCRIPTION 1-Channel T-Cube LED Driver (Requires TPS001) Power Supply for 1-Channel T-Cube LED Driver

Breakout Box for 4-Channel LED Drivers

Components*

(Steps 6 and 10 on Pages 22 - 23)

ITEM # WFA5100

PRICE 381.11

DESCRIPTION Single-Objective Prism Holder

WFA5110

199.56

WFA3000

206.67

MBN71950

1,268.00

MBD75300

1,346.00

Polarizer Turret

MBN71946

1,346.00

Visible Polarizer

MBN71960

2,021.00

NIR Polarizer

MBH72300

1,341.00

Dry DIC Condenser Prism, 10X

MBH72310

1,341.00

Dry DIC Condenser Prism, 16X - 100X

MBH76210

825.00

MBH76720

1,122.00

MBH76220

825.00

MBH76225

2,031.00

MBH76240

825.00

MBH76640

1,122.00

DIC Slider, 40X, for Plan Apo NIR (N2 Dry)

MBH76160

825.00

DIC Slider, 60X, for Plan Apo NIR (N2 Dry)

CFI175 to CFI160 Prism Adapter Turret Adapter Visible and NIR DIC Analyzer

DIC Slider, 10X, for Plan Fluorite DIC Slider, 16X DIC Slider, 20X, for Plan Fluorite or Plan Apo (N2 Dry) DIC Slider, 25X DIC Slider, 40X, for Plan Fluorite or Plan Apo (Air)

*Requires MBE74100 Nikon Epi-Illuminator, WFA100 DIC Module, and Condenser

Pricing for Pockels Cells, our Variable Attenuator, and our Variable Beam Expander is given on pages 149-151. 35

Imaging Systems Bergamo Series Microscopes

Overview: Cerna Series Microscopes

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

The Cerna Series is Thorlabs’ platform for electrophysiology, optogenetics, DNA injection, and other applications that require widefield and transmitted light imaging. Just like our Bergamo II Series for multiphoton microscopy, the Cerna Series adopts a modular approach to microscope construction that allows the user to configure the system to exact experimental requirements. The result is a versatile, expandable, accessible design that is fully compatible with Thorlabs-manufactured and user-supplied accessories.

Options at a Glance n Widefield

Imaging • Nikon Epi-Illuminator with 6-Position Turret • Thorlabs Single-Cube Epi-Illuminator • Nikon Trinoculars • High-Power LED or Liquid Light Guide Illumination Source • One or Two Scientific Cameras Dedicated to Transmitted Light or Fluorescence Detection n Transmitted Light Imaging • Nikon Condenser with 1" Motorized Travel • Differential Interference Contrast (DIC) • Dodt Contrast • Brightfield Imaging

n Sample

Holders with Motion Control • Manual or Motorized Micromanipulators for Patch Clamping • Fast XY Scanning Stage for Multiple Slides, Multiwell Plates, and Custom Inserts • Rigid Stands for Slides, Recording Chambers, and Custom Inserts, with Optional 1" Motorized Travel in X and Y • Low-Profile XY Platform with 2" Motorized Travel in X and Y n Objectives • Single Objective Holder, Dual Objective Changer, or 5-Position Objective Turret • 1" Motorized Travel in Z • Piezo Objective Holder for Fast Z-Stacks

Imaging Systems Bergamo Series Microscopes

Overview: Cerna Series Microscopes Minimal Footprint that Maximizes Available Working Space Cerna microscopes are engineered around Thorlabsâ&#x20AC;&#x2122; 95 mm optical construction rails. We chose these rails because they offer a linear dovetail mounting surface, excellent vibrational damping, and stable long-term support. These stiff rails were originally designed for users of our optomechanics who require rigid support for large, three-dimensional, load-bearing structures.

Cerna Series Microscopes Cerna Microscope Modules are Attached to a Rear-Mounted 95 mm Support Rail, Enabling LongTerm Mounting Stability with Clear Approach Angles

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy

The support rail is recessed with respect to the microscopeâ&#x20AC;&#x2122;s optical modules, leaving the approach angles to the objective unrestricted. Our open design is ideal for electrophysiology applications, as it provides access to the focal plane from the front, sides, and even behind. The rail also enables a vertical construction that consumes a minimal amount of space in the horizontal dimension, preserving valuable table space for experimental apparatuses.

Optical Tweezers

The standard height of the support rail is 400 mm. For setups that do not require transmitted light imaging, we offer a shorter 350 mm height that makes it easier to reach the trinoculars and objectives. The Compact Profile of the Cerna Microscope Body Leaves Ample Space for Experimental Equipment (Optical Table Size: 3' x 6')

Imaging Systems Bergamo Series Microscopes

Overview: Cerna Series Microscopes

Cerna Series Microscopes

User Expandable with Thorlabs, Industry-Standard, or User-Supplied Add-Ons

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

The modular design of Cerna accessories makes it simple to adapt these microscopes for a wide variety of imaging modalities. We manufacture several highperformance modules for these systems, including Dodt and differential interference contrast (DIC) modules for transmitted light imaging, scientific cameras for widefield and fluorescence imaging, and micromanipulators for patch clamping. Cerna microscopes are also directly compatible with several industry-standard components from Nikon, including their trinoculars, condensers, and epi-illuminator with 6-position dichroic turret. The experimental versatility of Cerna systems is aided by the linear dovetail mounting surface that spans the height of the 95 mm support rail. Modules such

as motorized condensers, sample holders, and our transmitted light imaging modules can be positioned along this surface using a balldriver. The dovetail centers the modules along the optical axis so that the user can avoid painstaking optical alignment procedures. For system developers who wish to build microscopes entirely from Thorlabsâ&#x20AC;&#x2122; catalog of parts (see page 43 for an example), the Cerna Series offers connections to our standard mechanical interfaces, including SM1 threads (1.035"-40), SM2 threads (2.035"-40), and taps for 30 mm and 60 mm cage compatibility. A breadboard accessory with either 1/4"-20 or M6 x 1.0 tapped holes attaches to the top of the support rail to allow the construction of custom illumination pathways.

Our Open Microscope Construction Allows the User to Modify and Optimize the System for Specific Experimental Needs

Imaging Systems Bergamo Series Microscopes

Overview: Cerna Series Microscopes

Cerna Series Microscopes

Compatible Accessories The image below highlights several components that are compatible with Cerna microscopes. This wide selection makes it easy to modify the system as experimental requirements evolve.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems

Objectives See Page 222

TDI Digital Microscopy

Filter Cubes and Filter Sets

Optical Tweezers

For Epi-Illumination

Scientific Cameras

See Pages 235 - 244

For Widefield or Transmitted Light Imaging See Page 308

LED Light Sources

For Brightfield Illumination, DIC Imaging, or Dodt Contrast See Page 484

Rigid Stands

For Slides, Recording Chambers, Z-Axis Piezo Stages, or Other Inserts See Page 296

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Sample System Configuration: Basic Cerna Microscope with DIC Imaging This Cerna microscope is configured for differential interference contrast (DIC) imaging. It incorporates several Nikon-designed DIC components, including their condensers, polarizers, and prisms. Illumination for the interference image is provided by Thorlabs’ high-power LEDs, which provide broadband illumination in the visible and NIR and are available at multiple wavelengths. By removing the DIC components from the optical path, these LEDs can be utilized for basic brightfield illumination. The Nikon epi-illuminator with slots for six dichroic filter cubes supports DIC imaging and enables epi-fluorescence studies. It is shown here equipped with a broad-spectrum LED lamp, but it can also accommodate an industrystandard liquid light guide or Thorlabs’ plasma light source (see page 514 for details).

Highlights n Nikon

Sliding Nosepiece Accommodates Two Objectives • Low-Magnification Objective for Finding the Region of Interest • High-Magnification Objective for High Resolution Imaging n 2D Stage Provides 2" Travel in X and Y for Microscope Body n Two Scientific Cameras can be Individually Dedicated to DIC Imaging and Widefield Fluorescence

Widefield Illumination • Broad-Spectrum LED Lamp • Nikon Epi-Illuminator with 6-Position Turret

Motion Control • 2" Travel in X and Y for Microscope Body

Sample Holder • Rigid Stand with Slide Holder

Transmitted Light Illumination • Brightfield and DIC Imaging • Visible and NIR Illumination • Nikon Condenser with 1" Travel

Widefield Viewing • Nikon Trinoculars • Two Dedicated Scientific Cameras

Imaging Systems

Sample System Configuration: Cerna Microscope for Electrophysiology In order to support electrophysiology, we added our motorized Burleigh micromanipulators (see pages 274 - 277) to the configuration on the previous page. These micromanipulators employ PZT piezoelectric elements for nanometer-resolution, backlash-free, low-drift positioning (<1 µm/hr of drift under temperature-controlled conditions). The micromanipulators and sample are secured on top of our rigid stands, which offer a quick-release clamp for ease of positioning.

Cerna Series Microscopes

Highlights n Recessed

95 mm Support Rail Leaves Approach Angles Open for Micromanipulators n Sample Holder is Motorized for 1" of Fine Travel in X and Y n Motorized Micromanipulators, Sample Holder, and Condenser Focusing Module Minimize the Chance of Unintended Movement

This setup is well suited for single-cell recordings, where it is not strictly necessary to move the micromanipulators and recording chamber together. For more complex setups, we manufacture an XY platform that provides a large motorized working surface (see the next page for more information).

Bergamo Series Microscopes

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Widefield Illumination • Liquid Light Guide • Nikon Epi-Illuminator with 6-Position Turret

Patch Clamping • Motorized Micromanipulators • Joystick Controller for X, Z, and Approach Axes • Knobbed Controller for Piezos

Widefield Viewing • Nikon Trinoculars • Two Dedicated Scientific Cameras

Sample Holder • Rigid Stand with Slide Holder • 1" Travel in X and Y

Transmitted Light Illumination • Brightfield and DIC Imaging • Visible and NIR Illumination • Nikon Condenser with 1" Travel

The image on the computer screen is courtesy of Dr. Lei Zhang and Professor Joshua Singer, University of Maryland.

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Sample System Configuration: Cerna Microscope for Electrophysiology and Confocal Microscopy Highlighting the modular construction of our Cerna microscopes, the system shown below has been configured for confocal imaging, while retaining the electrophysiology, DIC imaging, and epi-fluorescence imaging capabilities of the Cerna microscopes on the previous two pages. This fully functional confocal microscope uses Thorlabs’ galvoresonant scan head for raster scans of the field of view and our motorized pinhole wheel as the confocal aperture (each described in more detail on pages 58 - 59). In order to support electrophysiology setups, where it is critical to move the sample and the micromanipulators together, this configuration incorporates our XY platform. This low-profile platform allows micromanipulators, recording chambers, and other devices to be mounted in front of, alongside, or behind the objective, on a single common surface. It is controlled by our standard 3-axis controller with variable speed knobs.

Highlights n Motorized

XY Platform Moves Simple or Complex Apparatuses in Unison with the Sample • Large Working Surface: 24" x 18" (600 mm x 450 mm) • 2" Travel Range in X and Y • 1/4"-20 or M6 x 1.0 Tapped Holes for Securing Equipment n Scientific Camera and Nikon Trinoculars Visualize the Field of View n Fiber-Coupled Light Paths Greatly Simplify Tabletop Layout, Laser Alignment, and Maintenance

Widefield Viewing Widefield Illumination • Liquid Light Guide • Nikon Epi-Illuminator with 6-Position Turret

Patch Clamping • Motorized Micromanipulators • Joystick Controller for X, Z, and Approach Axes • Knobbed Controller for Piezos

Sample Holder

Transmitted Light Illumination • Brightfield and DIC Imaging • Visible and NIR Illumination • Nikon Condenser with 1" Travel

• XY Platform Moves Sample and Setup as One • 2" Travel in X and Y • 3-Axis Controller with Variable Speed Control

• Nikon Trinoculars • Nikon Epi-Illuminator with 6-Position Turret • Scientific Camera

Imaging Systems

Sample System Configuration: Developers’ Configuration The configuration below contains several microscope body attachments that are specifically designed for expanding the microscope’s functionality using Thorlabs’ mechanical construction systems (see pages 408 - 409 for an introduction). These accessories, intended for system developers, enable a high degree of system customization. For instance, the 95 mm support rail has been capped with a breadboard that has a Ø1.5" through hole and an array of 1/4"-20 taps, which here have been used to build a widefield viewing apparatus with our plasma light source and scientific camera. Taking advantage of the linear dovetail mounting surface on the support rail, we have attached an SM1-threaded (1.035"-40), 30-mm-cage-compatible fixed arm and an SM2-threaded (2.035"-40), 60-mm-cage-compatible fixed arm in the optical path underneath the objective, allowing us to construct custom transmitted light illumination apparatuses. Fixed Arms for Microscope Body • Hold Samples or Optics Along Dovetail Mounting Surface • Three Versions - SM1 Threads and Taps for 30 mm Cage Systems - SM2 Threads and Taps for 60 mm Cage Systems - Sized for MLS203-1 XY Scanning Stage (See Page 168)

Bergamo Series Microscopes Cerna Series Microscopes

Highlights n Fixed

Arms Integrate Thorlabs’ SM1, SM2, 30 mm Cage, and 60 mm Cage Construction Standards with the 95 mm Support Rail n Breadboard Top Enables User-Designed Widefield Viewing Schemes • Ø1.5" Through Hole for Optical Path • Array of 153 1/4"-20 or M6 x 1.0 Taps n Custom-Built Illumination Apparatuses Using Thorlabs’ Selection of Light Accessories • Fiber-Coupled Lasers (See Pages 480 - 483) • High-Power LEDs (See Pages 484 - 508) • Plasma Light Sources (See Page 514)

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Thorlabs is eager to assist with custom system builds, starting from either the catalog configurations shown here or parts from our components catalog. Please contact ImagingSales@thorlabs.com to start a conversation.

Breadboard Top • Used for Custom Epi-Illumination • 1/4"-20 or M6 x 1.0 Taps • Ø1.5" Through Hole

Illumination Sources • User Selects Wavelength and Output Power • Lamps, LEDs, and Lasers Available

Cerna Series: Electrophysiology Nikon MBH76210 DIC Slider 10X Plan Fluor

Nikon MBN71946 FN-P Visible Rotatable

Nikon MBH76640 DIC Slider 40X APO NIR (N2)

DIC Condenser Polarizers

Nikon MBH76720 DIC Slider 16X

Cerna Series Configuration C207 This configuration is designed for fast XY scans of slides, multiwell plates, and other inserts. It provides both epi-illumination and transmitted light illumination using high-power LEDs or liquid light guides.

Nikon MBH76240 DIC Slider 40X Plan Fluor, Plan APO (Air), Plan APO (LWD 40X 1.15 NA) Nikon MBH76220DIC Slider 20X Plan Fluor, Plan APO (N2 Dry)

Nikon MBH76225 DIC Slider 25X

Nikon MBH72300 N1 Dry (10X)

Nikon MBN71960 FN-IRP IR Rotatable

Nikon MBH72310 N2 Dry (16X - 100X)

DIC Condenser Prisms

Nikon MBD75300 FN-PT Polarizer Turret for VIS & IR Polarizers

DIC Objective Slider (Choose One Compatible w/ Your Objective)

Polarizers & Prisms for DIC Condenser

Nikon MBH76160 DIC Slider 60X APO NIR (N2)

DIC Analyzer

Nikon MBN71950 FN-IR/FA IR & VIS DIC Analyzer

WFA3000 DIC Turret Adapter

LED Drivers

DC4100 4 Ch 1 Mod Input

WFA1020 NIR LED

Choose DIC Components

Step 10: WFA0150 Transmitted Light Module Adapter

Illumination Kits

Choose a Body Height

Step 11:

WFA1000 Transmitted Light Only

WFA1010 VIS LED

Cerna Series Configuration C211 Equipped with a manual objective changer, two scientific cameras, and a Nikon epi-illuminator with 6-position turret, this configuration is optimized for widefield imaging under a variety of illumination conditions. (See Page 272 for Details)

LEDD1B 1 Ch

DC4104 4 Ch 4 Mod Input

Step 1:

Requires: • MBE74100 from Step 4 • Nikon Nosepiece from Step 5 • Brightfield Module from Step 10

WFA1100 Dodt/Gradient

Choose Transmitted Light Illumination

CSB1350 350 mm Tall Body

Brightfield Modules and Accessories

WFA1050 VIS & NIR LED

Step 9: Choose a Condenser

Step 8: Choose Motion Control BSA2000 Compact Condenser Mounting Arm

Nikon MBL70100 FN-C LWD 0.78 NA

CSA2000 Condenser Mounting Condenser Arm Mounting Arms

Nikon MBL78600 D-CUD Universal 0.9 NA

ZFM2020 Condenser Focus Module Nikon MBL78700 D-CUO DIC Oil 1.4 NA MCM3000 3-Axis Controller

Cerna Series Configuration C202 This configuration is ideal for patch clamping. It is centered around our XY platform, which offers a large, motorized workspace for moving the specimen and apparatus as one. (See Page 273 for Details)

PMP-2XY(/M) XY Manipulator Platform

PLS-X 1" X-Axis Stage MMP-2XY XY Mover for Microscope Body

PLS-XY 1" XY Stage

and Widefield Imaging Rigs MCWHL5 Cold White LED

1500M-GE Thorlabs Scientific Camera WFA4105 Nikon Y-TV Tube & C-Mount Adapter

WFA4100 1X Camera Port

WFA4106 Nikon Y-TV Tube & 0.7X C-Mount Adapter

WFA4101 0.75X Camera Port

WFA4000 Nikon Trinoculars w/ Eyepieces & IR Filter

WFA4102 0.5X Camera Port

With Trinoculars

CSA3000 Breadboard Top (Imperial) CSA3001 Breadboard Top (Metric)

Nikon MBD75100 FN-DP Variable Magnification Double Port

LEDs Illumination Sources

Nikon MBE74100 Epi-Illuminator w/ Turret

Not Required for Single Camera Configurations

M470L3 Blue (470 nm) LED Mounted High-Power LEDs (Only Compatible w/ WFA2001, Other Wavelengths are Available)

WFA2001 Epi-Illuminator

Without Trinoculars

Step 2:

M565L3 Lime (565 nm) LED

Nikon MBB74105 Y-IDP Dual Port

Lamps

Cerna Series Configuration C223 Our user-installable DIC imaging modules and a scientific camera have been attached to this microscope. Removing the modules allows the microscope to also function as a transmitted light stand.

X-CITE120LED High-Power, Broad-Spectrum LED (Only Compatible w/ MBE74100)

Step 3:

Choose Widefield Viewing Accessories

Choose a Double Camera Port

Epi-Illumination Module

X-CITE200 DC-Stabilized Fluorescence Light Source

HPLS243 Solid State Plasma Light Source

Step 4: Choose an Epi-Illuminator

CSA1100 Single Objective Arm

Step 5: Choose an Objective Holder

CSB1400 400 mm Tall Body

Step 6: Choose an Objective

N4X-PF

CSA1200 Sliding Nosepiece Arm

Nikon MBP71100 Nikon MBP70100 Single Objective FN-S2N Sliding Nosepiece 2-Position Nosepiece Nikon Nosepiece (Required for DIC in Step 11)

Step 7: Choose Sample/ Manipulator Stages

Rigid Stand Sample Holders

For Micromanipulators

For Slides

N10X-PF

N20X-PF

N40X-PF

N60X-PF

Other Objectives Available Upon Request Fixed Arms for Microscope Body

LOM1200 Manual 2-Position Nosepiece CSA1300 Thorlabs Sliding Nosepiece Arm

For Recording Chambers

MLS203-1 Fast XY Scanning Stage For MLS Inserts

Thorlabs Nosepiece

MOT1500 MOT2500 Manual 5- Motorized 5-Position Position Turret Turret (Coming Soon) CSA1400 Turret Mounting Arm

CSA1000 For MLS203-1 XY Stage

LOM2200 Motorized 2-Position Nosepiece

Cerna Series Configuration C218 This configuration highlights our rigid stand sample holder, which allows slides to be easily rotated into and out of the optical path. The rigid stand is mounted on motorized stages for 1" of travel in both X and Y.

CSA1001 SM1, 30 mm Cage Arm

Turret

CSA1002 SM2, 60 mm Cage Arm

MZS500-E Z-Axis Piezo Stage and Controller Kit

Cerna Series Configuration C229 This wholly customizable configuration is equipped with a breadboard top that allows users to build their own illumination pathways. Connections are provided for Thorlabsâ&#x20AC;&#x2122; SM1, SM2, 30 mm cage, and 60 mm cage construction standards. (See Page 43 for Details)

Imaging Systems Bergamo Series Microscopes

Cerna Configurations and Pricing

Cerna Series Microscopes

The labels on the sample Cerna system below correspond to the headings of the price boxes on the following three pages and are intended to help identify which price box contains the desired components. They also help identify the support modules required for each selected accessory.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Widefield Viewing Epi-Illumination • Choice of Epi-Illuminator or Breadboard Top - Thorlabs Single-Cube Epi-Illuminator - Nikon Epi-Illuminator with 6-Position Turret - Epi-Illuminators Require Filter Cubes and Filter Sets - Breadboard Tops: 1/4"-20 or M6 Taps • Choice of Illumination Source - Thorlabs’ High-Power LEDs - X-Cite Broad-Spectrum LED - X-Cite DC Lamp

• Trinoculars • Choice of Cameras - Basic CCD Camera - Thorlabs’ Scientific Cameras - Cameras Require Camera Tubes • Option: Double Camera Port

Nosepieces • Choice of Single Objective Holder, Dual Objective Nosepiece, or 5-Objective Turret • Requires Nosepiece Arm

Microscope Body • 350 mm or 400 mm Tall • Option: 2" Travel in X and Y

Transmitted Light Illumination • Choice of Condensers - Requires Mounting Arm and Focusing Module • Choice of DIC Imaging, Dodt Contrast, and/or Brightfield - Requires Illumination Kit and Driver - DIC Requires Nikon Epi-Illuminator and Nikon Nosepiece

Sample Holders Objectives

• Choice of Rigid Stands for Slides, Recording Chambers, or Inserts • Choice of Fixed Arms

• Choice of Objectives • Option: Piezo Objective Holder

Motion Controller

Motion Control Options

• 3-Axis Controller with Variable Speed Knobs • Required for Objective Focusing, Condenser Focusing, and Any X or Y Translation

• Rigid Stands: 1" Travel in X and/or Y • Microscope Body: 2" Travel in X and Y • XY Platform with 2" Travel in X and Y

Imaging Systems Bergamo Series Microscopes

Cerna Configurations and Pricing Microscope Body (Step 1 on Pages 44 - 45) ITEM # CSB1350 CSB1400

PRICE 3,542.63

DESCRIPTION 350 mm Tall Body

3,874.70

400 mm Tall Body

Widefield Viewing: Trinoculars (Step 2 on Pages 44 - 45) ITEM # WFA4000

PRICE 2,850.00

DESCRIPTION Nikon Trinoculars with Eyepieces

Widefield Viewing: Camera Tube (Step 2 on Pages 44 - 45) ITEM # WFA4100

PRICE 1,450.00

DESCRIPTION 1X Camera Tube for WFA2001 Single-Cube Epi-Illuminator

WFA4101

1,450.00

WFA4102

1,450.00

WFA4105

387.00

Nikon Y-TV Tube with C-Mount for MBE74100 Nikon Epi-Illuminator

WFA4106

764.00

Nikon Y-TV Tube with 0.7X C-Mount for MBE74100 Nikon Epi-Illuminator

0.75X Camera Tube for WFA2001 Single-Cube Epi-Illuminator 0.5X Camera Tube for WFA2001 Single-Cube Epi-Illuminator

Widefield Viewing: Cameras (Step 2 on Pages 44 - 45) ITEM # DCU224M

PRICE 2,184.00

DESCRIPTION Monochrome CCD Camera

340M-GE

4,400.00

Fast-Frame-Rate VGA Scientific Camera

1500M-GE

5,100.00

1.4 Megapixel Scientific Camera

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Widefield Viewing: Double Camera Ports* (Step 3 on Pages 44 - 45) ITEM # MBB74105

PRICE 2,010.00

MBD75100

8,519.00

DESCRIPTION Nikon Y-IDP Double Camera Port Module Nikon FN-DP Variable Magnification Double Camera Port Module

*Not Required for Single Camera Configurations

Epi-Illumination (Step 4 on Pages 44 - 45) ITEM # WFA2001

PRICE 2,300.00

MDFM-MF2

473.89

MBE74100

2,797.00

MDFM-TE2000

466.60

Epi-Illumination: Breadboard Top (Step 4 on Pages 44 - 45) ITEM # METRIC ITEM # CSA3000 CSA3001

PRICE 992.15

DESCRIPTION Breadboard Top, 1/4"-20 (M6) Taps

Epi-Illumination: Filter Set ITEM # MDF-BFP

PRICE 625.00

DESCRIPTION Blue Fluorescent Protein (BFP) Emission, Excitation, and Dichroic Filter Set

MDF-GFP2

745.00

Alexa FluorÂ® 488 Emission, Excitation, and Dichroic Filter Set

MDF-MCHB

745.00

mCherryB Emission, Excitation, and Dichroic Filter Set

MDF-TOM

745.00

tdTomato Emission, Excitation, and Dichroic Filter Set

Epi-Illumination: High-Power LEDs* (Step 4 on Pages 44 - 45) ITEM # M470L3

PRICE 260.00

DESCRIPTION 470 nm LED

M565L3

205.00

M940L3

205.00

940 nm LED

MCWHL5

187.51

Cold White, 6500 K LED

565 nm LED

*Driver Must be Purchased Separately

Epi-Illumination: High-Power LED Driver (Step 9 on Pages 44 - 45) ITEM # LEDD1B

PRICE 284.00

DESCRIPTION 1-Channel T-Cube LED Driver (Requires TPS001)

TPS001

25.00

DC4100

2,505.00

4-Channel LED Driver, 1 Modulation Input (Requires DC4100-HUB)

DC4104

2,759.00

4-Channel LED Driver, 4 Modulation Inputs (Requires DC4100-HUB)

DC4100-HUB

300.00

Power Supply for 1-Channel T-Cube LED Driver

Breakout Box for 4-Channel LED Drivers

Epi-Illumination: Other Illumination Sources (Step 4 on Pages 44 - 45) ITEM # HPLS243

PRICE 3,810.00

DESCRIPTION Plasma Light Source with Liquid Light Guide

X-CITE120LED

6,495.00

Broad-Spectrum LED Lamp

X-CITE 200

6,495.00

DC Lamp with Liquid Light Guide

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes

Cerna Configurations and Pricing Nosepieces: Single Objective (Step 5 on Pages 44 - 45) ITEM # CSA1100

PRICE 205.43

MBP71100

DESCRIPTION Single Objective Arm

666.00

Manual Dual Objective Nosepiece (Requires CSA1300)

Nosepieces: Dual Objective (Step 5 on Pages 44 - 45) ITEM # MBP70100

ThorImageLS Software

LOM1200

$ 3,681.11

Manual Dual Objective Nosepiece (Requires CSA1300)

LOM2200

$ 5,313.26

Motorized Dual Objective Nosepiece (Requires CSA1300)

OCT Systems

Nosepieces: 5-Objective Turret (Step 5 on Pages 44 - 45)

TDI Digital Microscopy Optical Tweezers

ITEM # MOT1500

PRICE 1,791.00

DESCRIPTION Nikon FN-S2N Sliding Dual Objective Nosepiece (Requires CSA1200)

PRICE $ 2,979.34

DESCRIPTION Manual 5-Objective Turret (Requires CSA1400)

Nosepieces: Arms (Step 5 on Pages 44 - 45) ITEM # CSA1200

PRICE 240.19

DESCRIPTION Nikon Nosepiece Arm

CSA1300

$ 301.77

Nosepiece Arm for LOM1200 or LOM2200

CSA1400

$ 293.20

Turret Arm for MOT1500 or MOT2500

Objectives (Step 6 on Pages 44 - 45)

Pricing for Our Micromanipulators is on Pages 275 and 277

ITEM # N4X-PF

PRICE 432.00

N10X-PF

785.00

N20X-PFH

6,706.00

N40X-PF

991.00

N60X-PF

2,334.00

DESCRIPTION Nikon Plan Fluorite Objective, 4X, 0.13 NA, 17.2 mm WD Nikon Plan Fluorite Objective, 10X, 0.3 NA, 16 mm WD Nikon Plan Fluorite Objective, 20X, 0.50 NA, 2.1 mm WD Nikon Plan Fluorite Objective, 40X, 0.75 NA, 0.66 mm WD Nikon Plan Fluorite Objective, 60X, 0.85 NA, 0.31 to 0.4 mm WD

Objectives: Piezo Objective Holders ITEM # PI-P725

PRICE 15,499.00

DESCRIPTION Physik Instrumente (PI) Long-Travel Objective Stage (Requires SC-IMG2)

PI-P726

15,090.00

Physik Instrumente (PI) Piezo Scanner for Heavy Objectives (Requires SC-IMG2)

SC-IMG2

1,500.00

Scan NI Package

Sample Holders: Fixed Arms for Microscope Body (Step 7 on Pages 44 - 45) ITEM # CSA1000

PRICE 979.49

DESCRIPTION Holder for MLS203-1 Fast XY Scanning Stage (See Page 168)

CSA1001

408.78

Arm with Ø1", SM1-Threaded Hole and Taps for 30 mm Cage Systems

CSA1002

412.63

Arm with Ø2", SM2-Threaded Hole and Taps for 60 mm Cage Systems

Sample Holders: Rigid Stands* (Step 7 on Pages 44 - 45) ITEM # MP100-RCH2

PRICE 752.91

DESCRIPTION Rigid Stand with Slide Holder, Adjustment Height: 148.1 - 208.5 mm

MP150-RCH2

766.24

Rigid Stand with Slide Holder, Adjustment Height: 198.1 - 309.3 mm

MP200-RCH2

779.58

Rigid Stand with Slide Holder, Adjustment Height: 248.1 - 410.1 mm

MP250-RCH2

792.91

Rigid Stand with Slide Holder, Adjustment Height: 265.9 - 510.9 mm

MP100-RCH1

648.22

Rigid Stand with Recording Chamber Holder, Adjustment Height: 148.1 - 208.5 mm

MP150-RCH1

661.56

Rigid Stand with Recording Chamber Holder, Adjustment Height: 198.1 - 309.3 mm

MP200-RCH1

674.89

Rigid Stand with Recording Chamber Holder, Adjustment Height: 248.1 - 410.1 mm

MP250-RCH1

688.22

Rigid Stand with Recording Chamber Holder, Adjustment Height: 265.9 - 510.9 mm

MP100-MLSH

769.84

Rigid Stand with Insert Holder, Adjustment Height: 148.1 - 208.5 mm

MP150-MLSH

783.18

Rigid Stand with Insert Holder, Adjustment Height: 198.1 - 309.3 mm

MP200-MLSH

796.51

Rigid Stand with Insert Holder, Adjustment Height: 248.1 - 410.1 mm

MP250-MLSH

809.84

Rigid Stand with Insert Holder, Adjustment Height: 265.9 - 510.9 mm

*See Pages 296 - 300 for Full Presentation

Motion Control Options (Step 8 on Pages 44 - 45)

ITEM # PLS-X

METRIC ITEM # –

PRICE 1,296.80

DESCRIPTION 1D Stage with 1" Travel in X for Rigid Stands (Requires MCM3000)

PLS-XY

–

2,370.52

2D Stage with 1" Travel in X and Y for Rigid Stands (Requires MCM3000)

MMP-2XY

–

12,865.00

2D Stage with 2" Travel in X and Y for Microscope Body (Requires MCM3000)

PMP-2XY

PMP-2XY/M

12,211.72

XY Platform with 2" Travel in X and Y and 1/4"-20 (M6) Taps (Requires Motion Controller)

Imaging Systems Bergamo Series Microscopes

Cerna Configurations and Pricing Motion Controller (Step 8 on Pages 44 - 45) ITEM # MCM3000

PRICE 4,500.00

DESCRIPTION 3-Axis Controller

Transmitted Light Illumination: Condensers (Step 9 on Pages 44 - 45) ITEM # MBL70100

PRICE 1,943.00

DESCRIPTION Nikon FN-C LWD Condenser, 0.78 NA

MBL78600

$ 1,836.00

Nikon D-CUD Condenser, 0.9 NA

MBL78700

$ 2,227.00

Nikon D-CUO DIC Oil Condenser, 1.4 NA

Transmitted Light Illumination: Condenser Focusing Module and Controller (Steps 8 and 9 on Pages 44 - 45) ITEM # ZFM2020

PRICE $ 1,687.50

DESCRIPTION Focusing Module with 1" Motorized Travel (Requires MCM3000)

MCM3000

$ 4,500.00

3-Axis Controller

Transmitted Light Illumination: Condenser Mounting Arm (Step 9 on Pages 44 - 45) ITEM # BSA2000

PRICE $ 850.00

DESCRIPTION Compact Condenser Mounting Arm with ±2.5 mm Travel in X and Y

CSA2000

$ 678.00

Condenser Mounting Arm with ±2 mm Travel in X and Y

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Transmitted Light Illumination: Brightfield Modules (Step 10 on Pages 44 - 45) ITEM # WFA0150

PRICE 337.60

DESCRIPTION Transmitted Light Module Adapter (Required for All Following Modules)

WFA1000

4,785.78

DIC Imaging Module (Requires Illumination Kit)

WFA1100

5,008.00

Dodt Contrast Module (Requires Illumination Kit)

Transmitted Light Illumination: Illumination Kits* (Step 9 on Pages 44 - 45) ITEM # WFA1010

PRICE $ 750.83

DESCRIPTION Visible LED Kit

WFA1020

$ 768.83

NIR LED Kit

WFA1050

$ 1,308.13

Visible and NIR LED Kit

*Driver Must be Purchased Separately

Transmitted Light Illumination: Illumination Kit Driver (Step 9 on Pages 44 - 45) ITEM # LEDD1B

PRICE 284.00

DESCRIPTION 1-Channel T-Cube LED Driver (Requires TPS001)

TPS001

25.00

DC4100

2,505.00

4-Channel LED Driver, 1 Modulation Input (Requires DC4100-HUB)

DC4104

2,759.00

4-Channel LED Driver, 4 Modulation Inputs (Requires DC4100-HUB)

DC4100-HUB

300.00

Power Supply for 1-Channel T-Cube LED Driver

Breakout Box for 4-Channel LED Drivers

Transmitted Light Modules: DIC Components* (Step 11 on Pages 44 - 45) ITEM # WFA3000

PRICE 206.67

DESCRIPTION Turret Adapter

MBN71950

1,268.00

MBD75300

1,346.00

Polarizer Turret

MBN71946

1,346.00

Visible Polarizer

MBN71960

2,021.00

NIR Polarizer

MBH72300

1,341.00

Dry DIC Condenser Prism, 10X

MBH72310

1,341.00

Dry DIC Condenser Prism, 16X - 100X

MBH76210

825.00

MBH76720

1,122.00

MBH76220

825.00

MBH76225

2,031.00

MBH76240

825.00

MBH76640

1,122.00

DIC Slider, 40X, for Plan Apo NIR (N2 Dry)

MBH76160

825.00

DIC Slider, 60X, for Plan Apo NIR (N2 Dry)

Visible and NIR DIC Analyzer

DIC Slider, 10X, for Plan Fluorite DIC Slider, 16X DIC Slider, 20X, for Plan Fluorite or Plan Apo (N2 Dry) DIC Slider, 25X DIC Slider, 40X, for Plan Fluorite or Plan Apo (Air)

*Requires MBE74100 Nikon Epi-Illuminator, MBP71100 or MBP70100 Nikon Nosepiece, and WFA1000 DIC Module

Imaging Systems Bergamo Series Microscopes

Essentials Kit Optimized Optical Scan Path, Ready for a Laser and Objective

Cerna Series Microscopes Essentials Kit Confocal Microscopes

Galvo-Resonant Scan Head and Two-Channel Detection Module

ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

ThorImageLS™ Software with Computer Electronic Control Unit

Thorlabs’ Multiphoton Essentials Kit helps customers build their own multiphoton microscopes. The core of the kit is a galvo-resonant scan path designed and manufactured by Thorlabs, ready to accept a femtosecond laser and multiphoton objective. Consisting of a galvo-resonant scan head, matched scan and tube lenses, and two GaAsP PMT detection channels in a single enclosure that maintains the optical alignment, this kit incorporates many of the same technologies and components used by our original Bergamo Series Multiphoton Microscopes. The spine of the system is Thorlabs’ 66 mm construction rail, which provides a vertical mounting surface for the optical system. Since the rail is symmetric, the scan path can be mounted at any height. This feature provides a large, user-adjustable volume underneath the objective that is ideal for large samples and complex experimental apparatuses, which are frequently incompatible with typical sample stage distances. The compact size of the enclosure also minimizes the physical footprint occupied on the tabletop. 50

Features n For

Customers Building Their Own Multiphoton Microscopes n Ideal for Studying Thin Samples In Vitro n Height is Coarsely Adjustable to Accommodate Large Samples and Apparatuses n Video Frame Rate of 30 fps at 512 x 512 Pixels n Maximum Frame Rate of 400 fps at 512 x 32 Pixels n Two PMT Detection Channels with Easy-to-Reach Filters

Galvo-Resonant Scan Path The 8 kHz galvo-resonant scan head included with the essentials kit is the same component used in our multiphoton microscopes. The scan speed depends upon the pixel dimensions of the desired image. At 512 x 512 pixels, video frame rates of 30 fps are supported, while at 512 x 32 pixels, the maximum frame rate of 400 fps is obtained.

Imaging Systems Bergamo Series Microscopes

Essentials Kit Laser Source

Included Components n Thorlabs’

Galvo-Resonant Scan Head n 8 kHz Resonant Scanner n Two GaAsP PMTs with High Quantum Efficiency n ThorImageLS™ Software for Image Acquisition, Visualization, and Analysis n Dichroic and Emission Filter Holder n All Necessary Computer and Electronics Hardware and Cables

Cerna Series Microscopes Essentials Kit Confocal Microscopes

Galvo Mirror

Resonant Mirror Scan Lens

ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

We recommend pairing this galvo-resonant scan head with a Pockels cell (pages 454 - 455 contain our selection). Pockels cells help minimize laser exposure and prevent photodamage to sensitive samples by enabling edge blanking and high-speed masking. Overexposure can occur with any resonant scanner because the scanner’s pixel dwell time is longer at the left and right edges of the scan than in the middle; edge blanking effectively blocks the laser when it is pointing at the edges of the field of view. High-speed masking further prevents photodamage by limiting the exposure to user-selected regions within the field of view.

PMT 1

Tube Lens

Emission Filter 1

PMT 2 Emission Filter 2

Primary Dichoric

Emission Dichoric

2-Channel PMT Detection Module Two-channel detection is provided by high-sensitivity, cooled GaAsP PMTs. The high quantum efficiency of these detectors helps capture signals from weakly fluorescent samples and also prevents photobleaching by allowing the use of lower laser powers. In order to protect the detector, these PMTs are designed to trip and turn off if the sensor becomes saturated.

Objective

Optical System Construction and Beam Path 51

Imaging Systems Bergamo Series Microscopes

Essentials Kit

Cerna Series Microscopes

Easy-to-Reach Dichroic and Emission Filter Holder

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

The primary dichroic and fluorescence emission filters are secured into the scan path by a holder that is specifically designed to maintain optical alignment. Access to this holder is available from the front of the enclosure. In order to optimize the system for different fluorescent tags, the user simply loosens a few screws; this process takes less than 5 minutes to complete. Our systems are fully compatible with industrystandard dichroics and emission filters. For our selection of these optics, please see pages 226 - 240.

External Trigger Support The electronic control unit (ECU) pictured on page 50 is the interface between the galvo-resonant scan head, the PMT detection channels, and the computer running ThorImageLS. It also contains inputs for electrical trigger signals, which can be used to synchronize experimental equipment.

ThorImageLS Capture Setup Tab

Triggers sent into the ECU can be used to start the acquisition of a single image or a series of images. The ECU can also be configured to output a trigger signal when the galvo-resonant scanner starts a new line or frame.

ThorImageLSâ&#x201E;˘ Software The multiphoton essentials kit includes the same software that powers our multiphoton microscopes. ThorImageLSâ&#x201E;˘ is a complete package for acquiring, visualizing, and analyzing multiphoton images. Its workflow-oriented interface supports single image, Z-stacks, time series, and image streaming acquisitions, and shows the user the measured images in real time. We provide a full SDK for our multiphoton systems that supports LabVIEW and C++. This SDK is utilized by the open-source ScanImage software suite by Vidrio Technologies (www.VidrioTechnologies.com), which is also compatible with the essentials kit.

Imaging Systems Bergamo Series Microscopes

Essentials Kit FEATURES AND BENEFITS OF THE MULTIPHOTON ESSENTIALS KIT

Excitation

Galvo-Resonant Scan Path

• High-Frame-Rate Functional Imaging - 30 FPS at 512 x 512 Pixels - 400 FPS at 512 x 32 Pixels

Two-Channel PMT Detection Module

• Full-Field-of-View Collection Optics Matched to Back Aperture of Popular Multiphoton Objectives • GaAsP PMTs with High Quantum Efficiency

Choice of ThorImageLS™, ScanImage, or Your Own Software

• ThorImageLS: Our Internally Developed Solution • ScanImage: 4.x or 5.x • Full SDK for LabVIEW and C++

Easy-to-Reach Dichroic and Emission Filter Holder

Input and Output Triggers

Large, Adjustable Volume Underneath Objective

Wavelength Range

• Filters are Accessed from the Front of the Scan Path and Take Less than 5 Minutes to Exchange • Use Electrical Signals to Synchronize All Your Equipment • Input Triggers can Start a Single Series or an Indefinite Series • Output Triggers can be Sent at the Beginning of a Frame or Line

PRICE 84,000.00

MPM-SCAN4

50,000.00

Field of View Microscope Objective Thread Scanners

680 - 1400 nm 16 mm Diagonal Square (Max) at the Intermediate Image Plane M32 See Page 405 for Adapters for RMS or M25 Objectives 8 kHz Resonant Scanner (X) Galvanometric Scan Mirror (Y)

Essentials Kit Confocal Microscopes ThorImageLS Software

Scan Speed

30 fps at 512 x 512 Pixels 400 fps at 512 x 32 Pixels

OCT Systems

Scan Mode

Line, Square, or Rectangle

Scan Zoom

1X to 36X

TDI Digital Microscopy

Scan Resolution

Up to 2048 x 2048 Pixels (Bi-Directional) Up to 4096 x 4096 Pixels (Unidirectional)

Optical Tweezers

Detection Backward Direction

• Accommodates Large Preps and Setups • Easily Adapt the Microscope for Differently Sized Experimental Setups

ITEM # MPM-2PKIT

Cerna Series Microscopes

MULTIPHOTON ESSENTIALS KIT

Two Ultrasensitive GaAsP PMTs

Collection Optics

Full- or Extended-Field-of-View User-Changeable Emission Filters and Dichroic

Wavelength Range

300 - 720 nm

DESCRIPTION Multiphoton Essentials Kit: Galvo-Resonant Scan Head, 2 PMTs, ECU, and Data Acquisition Computer Galvo-Resonant Scan Head, ECU, and Data Acquisiton Computer

Have you seen our...

Multiphoton Physiology Objectives

Fluorescence Imaging Filters

The full-field-of-view collection optics used in the Multiphoton Essentials Kit are optimized to match the back aperture of multiphoton objectives. Thorlabs stocks a selection of popular options from Nikon and Olympus.

Thorlabs’ Emission Filters and Dichroic Beamsplitters are designed for full compatibility with the Multiphoton Essentials Kit. Our emission filters are tuned for use with many common fluorophore types, premounted for ease of use, and labeled to aid in identification. Our selection of dichroic beamsplitters encompasses 15 different dichroic coatings, making it easy to find a beamsplitter that optimizes your system’s measurement sensitivity.

N60X-NIR

DMLP490R MDF-FITC

N16XLWD-PF

See Page 222

N20X-PFH

DMLP605R

DMSP1500R

See Pages 226 - 245

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System Thorlabs’ Confocal Microscopy System installed on our Cerna microscope (see pages 270 - 273). Several accessories have been added to the system, including Patch-Clamp Manipulators (see pages 274 - 279) and the PMP-2XY Motorized Platform (see pages 294 - 295).

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Features n Convert

a Standard Research-Grade Microscope into a Confocal Imaging System n Galvo-Resonant Scan Head • Video Rate Image Acquisition: 30 fps at 512 x 512 Pixels • 400 fps at 512 x 32 Pixels n Maximum Scan Resolution • 2048 x 2048 Pixels (Bi-Directional) • 4096 x 4096 Pixels (Uni-Directional) n Thorlabs-Designed Scan Lens Corrected for Chromatic Aberrations from 405 to 1100 nm

* See facing page for a summary of laser wavelength options and included pinhole sizes.

Thorlabs’ Confocal Laser Scanning Microscopy Systems allow researchers to add confocal imaging capabilities to their research-grade microscopes. The modular components are all developed and built in-house to maximize ease of use, simplify integration with existing imaging setups, and produce high-quality images. These systems offer turnkey integration with virtually any upright or inverted microscope (not included), provided 54

n Laser

Source with up to 4 Excitation Wavelengths* • All Sources Contain a Blue Laser for 488 nm • Wavelength Combinations are Optimized for Popular Fluorophores n Configurable Detector Module • Select 2 or 4 Detection Channels • Standard Multialkali or High-Sensitivity GaAsP PMTs • Filters Mounted in Kinematic Filter Cubes for Easy Exchange n 16-Position Motorized Pinhole Wheel with Round Pinholes* n ThorImageLS™ Software Suite for Microscope Control, Automated Data Collection, and Image Review that there is access to the intermediate image plane (e.g., camera port) via a C-mount. The systems can be controlled using the included ThorImageLS™ software, which features an intuitive graphical user interface and provides a means for quick data recording and review. These powerful capabilities work in concert to ensure that your equipment never constrains the quality of your results.

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System Our Confocal system features a galvo-resonant scan head that can capture up to 400 fps at 512 x 32 pixels. The high frame rates support rapid imaging of live systems and minimize the effects of phototoxicity and unwanted photobleaching. Scan resolutions of up to 4096 x 4096 pixels (uni-directional) allow the system to produce high-resolution images while taking advantage of the large-field-of-view scan lens. The laser module includes up to four sources with excitation wavelengths from 405 nm to 660 nm to enable imaging of a wide range of fluorescent dyes. The laser source and the filter set included in the detection module are optimized for the excitation and emission

wavelengths of popular fluorophores (see pages 61 and 63 for more information). The detection module can be configured with two or four detection channels to enable multi-spectral imaging. Select standard multialkali photomultiplier tubes (PMTs) for samples with large dynamic range or GaAsP high-sensitivity PMTs to support imaging of weakly fluorescent samples. Our applications engineers are available to discuss the optimal configuration for your imaging 980 nm needs and can be contacted at 973-300-3000.

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Laser Source Wavelength Options 660 nm 642 nm 588 nm 561 nm 532 nm 488 nm 405 nm

Pinhole Sizes • Ø25 µm • Ø30 µm • Ø35 µm • Ø40 µm • Ø45 µm • Ø50 µm • Ø60 µm • Ø70 µm

• Ø80 µm • Ø90 µm • Ø100 µm • Ø125 µm • Ø200 µm • Ø300 µm • Ø1 mm • Ø2 mm

Thorlabs’ confocal system scan head is shown mounted to the side camera port of a Nikon Eclipse Ti-U inverted microscope. The MFC1 Motorized Focus Controller is located immediately to the right of the scan head. An MZS500-E Z-Axis Piezo Stage has been mounted on an MLS203-1 High-Speed XY Stage to enable motorized XYZ scanning.

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit

4-Channel Confocal Microscopy System: Configuration Options A 4-Channel Confocal System (See the Next Page for Configuration Options)

Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

5 2

3 7 6 4

Peach Worm The confocal system can be configured for Z-stacks by adding the MLS203-1 Fast XY Scanning Stage and the MZS500-E Z-Axis Piezo Stage. A volumetric projection generated from confocal images of autofluorescence of an insect is shown to the right. The image shows part of its abdomen and leg. • Imaged Area: 330 µm x 330 µm, Depth: 80 µm • Excitation Wavelength: 488 nm • Emission (Detection) Wavelength: 500 - 550 nm • Objective Lens: 60X Water Immersion, NA 1.0 56

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System: Configuration Options 1 Confocal Scan Head A High-Speed Galvo-Resonant Scanner Designed for Visible (SCAN-VIS) or UV+Visible (SCAN-UV) Wavelengths

Motorized Pinhole Wheel with Round Pinholes

Scan Lens

Cerna Series Microscopes

Essentials Kit

Confocal Microscopes ThorImageLS Software

980 nm

OCT Systems TDI Digital Microscopy

Wavelength Options

2 Laser Source • 2 to 4 Laser Lines • 4 88 nm (Blue) Source Standard in All Systems • Rigidly Coupled or Pigtailed to Output to Eliminate the Need for Realignment

Optical Tweezers

660 nm 642 nm 588 nm 561 nm 532 nm 488 nm 405 nm

3 Dichroic Filter Sets • Configured for Popular Fluorophore Combinations • Housed in Thorlabs’ DFM Dichroic Filter Mount to Enable Quick and Easy Filter Changes

Filters are Mounted in DFM Dichroic Filter Mounts for Easy Exchange

4 Photomultiplier Tubes

•C hoose a 2-Channel or 4-Channel Detector Configuration • Standard or High-Sensitivity PMTs CLSD-2SS

CLSD-4HS

2-Channel Detection with Standard PMTs

4-Channel Detection with High-Sensitivity PMTs

Optional Accessories (Available Separately) 5 MLS203-1 Fast XY Scanning Stage and BBD202 Brushless Motor Controller 6 MJC001 2-Axis Microscopy Joystick Controller 7 MFC1 Motorized Microscope Focus Controller Not Shown in the Setup: • MZS500-E Z-Axis Piezo Stage and Controller Kit • MZF001 Z-Axis Joystick

MZF001 Z-Axis Joystick

The MZS500-E Z-Axis Piezo Stage Shown Installed in the MLS203-1 Fast XY Scanning Stage. An MZS500P2 Slide Holder is Mounted Inside the Z-Axis Stage.

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System

Cerna Series Microscopes

Confocal Scan Head

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Confocal Scan Head Mounted on a Cerna Microscope

Our confocal scan head consists of three principal components: a high-speed resonant scanner, a widefield-of-view scan lens, and a pinhole wheel output. The scanner can be attached to virtually any upright or inverted microscope with access to the intermediate image plane via a C-mount-threaded port (e.g., a camera port) so long as 100% of the output can be directed to this port. Light enters the scan head through one of the laser inputs, is reflected onto the galvo-resonant scanner mirrors by the primary filter block, and exits the assembly through the wide-field-of-view scan lens. Light then enters the microscope and excites fluorophores in the sample. The resulting fluorescence travels back through the scan lens and is reflected by the galvoresonant scanner to the pinhole wheel. The pinhole wheel output is connected to the PMT detection module via an SMA-terminated fiber.

High Speed Resonant Scanner Resonant scanners offer the advantage of highspeed image acquisition (up to 400 fps) and lower phototoxicity than slower scanning methods. Using Visible Laser Input

advanced digitization algorithms proprietary to Thorlabs, we have overcome the limitations imposed by the non-constant dwell time associated with sinusoidal scanning. The algorithm also allows us to take advantage of more of the drive waveform to provide the largest field of view for a resonant scanner system and makes every photon collected for each pixel count.

UV Laser Input

Visible Laser Input

Resonant Scanner

UV Laser Input

Pinhole Wheel Scan Lens

To Microscope

Pinhole Output to PMT

Galvo Scanner

Scan Lens

Primary Dichroic Beamsplitter

To Microscope 58

To PMTs

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System Pinhole Shape Comparison

Wide-Field-of-View Scan Lens To complement the large angular range over which the resonant scanner is used, Thorlabs’ engineers developed a scan lens optimized for large fields of view. The lens features excellent chromatic aberration correction from 405 to 1100 nm, superb field flatness, and very low distortion.

Airy Disk

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems

Motorized Pinhole Wheel The motorized pinhole wheel allows the pinhole size to be adjusted for a variety of imaging configurations and objective numerical apertures in order to simultaneously maximize the in-focus light that reaches the PMT detectors and minimize the transmission of signal from outside the focal plane. A rotating glass plate has 16 sizes of lithographic round pinholes deposited to exceedingly tight tolerances, ensuring that optimal alignment is maintained as each pinhole is rotated into the light path.

Cerna Series Microscopes

Cut-Away View of the Scan Lens

Airy Disk Seen Through a Round Pinhole

TDI Digital Microscopy Optical Tweezers

A round pinhole matches the shape of the point spread function, cleanly isolating the in-focus light while blocking out-of-focus light.

Motorized Pinhole Wheel Shown with the SMA Output Port Removed. (See Page 160 for More Details)

For thicker samples, the size of the pinhole should be optimized relative to the NA of the objective in order to maximize signal to noise. With Pinhole Sizes this in mind, our engineers selected each • Ø25 µm • Ø80 µm pinhole size in the motorized pinhole • Ø30 µm • Ø90 µm wheel to complement a common • Ø35 µm • Ø100 µm objective NA. Conversely, for thinner • Ø40 µm • Ø125 µm samples that produce less light outside • Ø45 µm • Ø200 µm of the focal plane, a larger pinhole size • Ø50 µm • Ø300 µm can help improve throughput. Pinhole • Ø60 µm • Ø1 mm diameters up to 2 mm provide flexibility • Ø70 µm • Ø2 mm so that you can easily adapt the system to your experiment. The diagram to the right illustrates the effects of several common pinhole shapes on the signal that reaches your detector. A round pinhole is the ideal shape for maximizing the transmission of light generated in the focal plane of your sample while also optimizing the rejection of signal generated above and below the layer that is being scanned.

Airy Disk Seen Through a Large Square Pinhole

A large square pinhole superscribed on the Airy Disk allows all of the in-focus light to pass, but some of the out-of-focus light will leak through as well.

Airy Disk Seen Through a Small Square Pinhole

A small square pinhole inscribed on the peak of the Airy Disk blocks out-of-focus light but also eliminates some of the usable signal.

980 nm

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System

Cerna Series Microscopes

Integrated Laser Source

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Wavelength Options

The solid state multilaser 660 nm 642 nm source minimizes maintenance 588 nm with an all-fiber design. Each 561 nm 532 nm laser line is individually fiber 488 nm coupled using a permanent rigid system. The individual 405 nm fiber-coupled lasers are then Laser Source combined in an all-fiber coupler. This design ensures that the lasers never go out of alignment, keeping the full power of the lasers coupled into the fiber at all times. For added flexibility, a second fiber output is provided that is dedicated to an optional 405 nm laser diode.

Expandable Detection Module Future-proof your experiments with our remotely positioned detector module that can be readily expanded from two to four photomultiplier tubes (PMTs). Sitting in front of each PMT is a quickly exchangeable dichroic beamsplitter and emission filter holder. The detector module can be configured with our standard-sensitivity multialkali PMTs or high-sensitivity GaAsP PMTs.

Detection Module

Filters Each confocal system includes an appropriate set of emission filters that block laser light from entering the PMTs and provide pass bands at the fluorescence wavelengths of popular fluorophores (see pages 226 - 231 for more filter options). The exact configuration is determined by the laser wavelengths in your confocal system. A common configuration with compatible fluorophores is outlined on the next page as an example. If you have questions concerning the filter set included with a specific laser configuration, please contact ImagingSales@thorlabs.com for more information.

Imaging Filter Set

Hippocampal Neurons Hippocampal culture stained with MAP2 and synapsin I antibodies.

Live hippocampal neurons stained with Lucifer Yellow.

Courtesy of Elena Popugaevu, St. Petersburg State Polytechnical University, St. Petersburg, Russia

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System

Cerna Series Microscopes

Sample Configuration The primary dichroic and emission filter sets in a confocal system are typically optimized for one of two excitation wavelength combinations. The most popular configuration is compatible with 405 nm, 488 nm, 561 nm, and 642 nm excitation lasers. The graph below provides and example of this common emission filter configuration, with the emission spectra of four compatible fluorophores superimposed on the filtersâ&#x20AC;&#x2122; transmission profiles. The second most common 4-laser configuration exchanges the 532 nm laser for a 561 nm laser, useful Configuration for Source with for samples thatFilter are marked with TRITC. To

accommodate this wavelength, the two bandpass filters centered at 525 nm and 593 nm would be replaced with a narrower bandpass filter (25 nm pass bandwidth) centered at 512 nm and a wider bandpass filter (75 nm pass bandwidth) centered at 582 nm. Other wavelength combinations are available upon request. Our team of applications engineers can help you determine the optimal excitation laser and emission filter configuration for your research. Please contact ImagingSales@thorlabs.com for more information.

Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

405 nm, 488 nm, 561 nm, and 642 nm Lasers Filter Configuration for Source with 405 nm, 488 nm, 561 nm, and 642 nm Lasers

100 100

Filters

Transmission (%) Transmission (%)

Essentials Kit

CWL: 447 nm/Pass Bandwidth: 60 nm (T avg > 93%) Filters CWL: 525 nm/Pass Bandwidth: 50 nm (Tavg > 93%) CWL: 593 nm/Pass nm/PassBandwidth: Bandwidth:6046nm nm(T(T 94%) avg > >93%) CWL: 447 avg 660 upnm to 1200 avg > 93%50 CWL:nm 525Longpass nm/Pass (T Bandwidth: (Tavg >nm) 93%) CWL: 593 nm/Pass Bandwidth: 46 nm (Tavg > 94%) Fluorophores 660 nm Longpass (Tavg > 93% up to 1200 nm) DAPI Fluorophores EGFP DAPI tdTomato EGFP Cy5 tdTomato The solid lines inCy5 the plot show the pass bands of

60 60

40 40

20 20

the filters, and the shaded areas show the normalized emission spectrum for the fluorophores.

0 350 0

400 350

450 400

500 450

550

600

650

500 550 600 Wavelength (nm) 650 Wavelength (nm)

700 700

750 750

800 800

EXCITATION Laser Source Laser Wavelength

405 nm

488 nm

561 nm

642 nm

EMISSION Emission Filters (Included Filters Determined by Laser Configuration)

Bandpass (CWL/Pass Bandwidth, Tavg)

447 nm/60 nm, >93% (Only with 405 nm Laser in Source) 525 nm/50 nm, >93% 593 nm/46 nm, >94%

Longpass

660 nm, Tavg >93% (up to 1200 nm)

Confocal Scan of C. elegans C. elegans motor neurons and muscle arms. The figure above shows the C. elegans strain, trIs30, expressing YFP in body wall muscles (green) and DsRED2 in the ventral nerve cord and motor neurons (red). Courtesy of Dr. William Ryu, University of Toronto

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System

Cerna Series Microscopes

ThorImageLS Data Acquisition Software

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Our ThorImageLS Software Package was developed in conjunction with our Confocal and Multiphoton Microscopy Systems to provide a seamless, logical, and intuitive program for acquiring and analyzing images. The workflow-oriented interface only displays the parameters you need for each scan series (such as Z series for volumetric scans or time series for imaging of dynamics). The software is designed to streamline the image acquisition process by dividing it into three steps: adjusting the settings of your imaging rig, acquiring the images, and data review and analysis. Each of these functions is controlled from a separate tab within the software with dropdown menus for the features that you need for your experiment. A fourth tab is provided for scripting so that more advanced users can automate their experiments.

Features n Independently

Control the Power of up to 4 Excitation Lasers n Select 1 of 16 Pinhole Diameters n Select Scan Area and Image Pixel Size n Assign a Color to Each Detection Channel n Generate 3D Z-Stack Reconstructions n Built-In Movie Maker: Animate Z-Stacks or Time Series to Share your Results n Data Saved in Lossless OME-TIFF Format â&#x20AC;˘ Compatible with ImageJ and Other Popular Image Analysis Programs â&#x20AC;˘ Metadata Included for Correct Image Scaling in ImageJ

For an overview of the software that outlines features incorporated for both Confocal and Multiphoton imaging applications, see pages 66 - 69.

This panel allows the power of up to 4 laser sources to be set independently.

The ThorImageLS Capture Setup tab controls the settings of each component in the Confocal System. Expandable menus on the left sidebar allow parameters such as the laser power, scan area, and pinhole size to be adjusted before beginning the image acquisition process.

ThorImageLS uses the microscope objective magnification (set by the user) to calculate the pixel size of your scan area. Square, rectangular, or linear scan regions can be selected.

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System

Cerna Series Microscopes

Specifications The Confocal System can be configured with one of the laser sources listed below. Each laser source comes with a set of emission filters for the detection modules that complements the excitation wavelengths of popular fluorophores. Our applications engineers can help you select a combination suited to your experimental needs. Contact ImagingSales@thorlabs.com for more information.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Thorlabs’ Confocal System Installed on a Nikon FN1 Microscope

EXCITATION WAVELENGTHS

INCLUDED EMISSION FILTERS

BLUE

GREEN/ ORANGE

CMLS-A

–

488 nm

–

642 nm

525 nm/45 nm and 635 nm/Longpass

562 nm

CMLS-Bb

405 nm

488 nm

–

642 nm

447 nm/60 nm, 512 nm/25 nm, and 635 nm/Longpass

495 nm and 538 nm

CMLS-C

–

488 nm

532 nm

642 nm

513 nm/17 nm, 582 nm/75 nm, and 635 nm/Longpass

538 nm and 649 nm

CMLS-D

–

488 nm

561 nm

642 nm

525 nm/45 nm, 607/36 nm, and 635 nm/Longpass

562 nm and 649 nm

CMLS-Eb

405 nm

488 nm

532 nm

642 nm

447 nm/60 nm, 513 nm/17 nm, 582 nm/75 nm, and 635 nm/Longpass

495 nm, 538 nm, and 649 nm

CMLS-Fb

405 nm

488 nm

561 nm

642 nm

447 nm/60 nm, 525 nm/45 nm, 607 nm/36 nm, and 635 nm/Longpass

495 nm, 562 nm, and 649 nm

CMLS-Gb

405 nm

488 nm

588 nm

642 nm

447 nm/60 nm, 525 nm/45 nm, 615 nm/24 nm, and 635 nm/Longpass

495 nm, 605 nm, and 649 nm

CMLS-H

–

488 nm

–

660 nm

525 nm/39 nm and 697 nm/58 nm

562 nm

CMLS-I

–

488 nm

–

525 nm/45 nm

N/A

CMLS-J

–

488 nm

532 nm

–

512 nm/25 nm and 582 nm/75 nm

538 nm

LASER SOURCE ITEM #a

RED

EMISSION FILTERS (CENTER WAVELENGTH/BANDWIDTH)

LONGPASS DICHROIC CUTOFF WAVELENGTH(S)

aFor

sources with less than four lasers, slots will be filled from left to right. bThese sources are only compatible with the SCAN-UV scan head.

Specifications for the galvo-resonant scanner included in each confocal system are listed below. These specifications are common to all of the confocal systems. See the following two pages for more information on configuration options. SCANNING Scan Head Galvo-Resonant Scan Speed Scan Zoom Scan Resolution

Galvo-Resonant Scan Head with 8 kHz Resonant Scanner (X) & Galvo Scan Mirror (Y) 30 fps at 512 x 512 Pixels 400 fps at 512 x 32 Pixels 2 fps at 4096 x 4096 Pixels 1X to 36X Bi-Directional: Up to 2048 x 2048 Pixels Unidirectional: Up to 4096 x 4096 Pixels

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System

Cerna Series Microscopes

Confocal System Compatibility Chart

Essentials Kit Confocal Microscopes ThorImageLS Software

The confocal system consists of four main modules: the scan head, laser source, detection module, and a computer. The chart below outlines the compatibility between these modules. Start by picking a scan head in the top row and follow the white lines down to see which other components are compatible. Thorlabs’ team of application engineers is also available to help you choose a system that is configured to suit your research needs. They can also help you determine if you will need any additional components, such as specialized camera tubes, to optimize the system’s performance on your microscope.

OCT Systems TDI Digital Microscopy Optical Tweezers

Scan Head (Choose 1 )

Laser Source (Choose 1 Item #)

PMT Module (Choose 1 Item #)

SCAN-UV

SCAN-VIS

Scan Head with 2 Fiber Ports

Scan Head with 1 Fiber Port

ITEM #

WAVELENGTHS

ITEM #

CMLS-B

405 nm, 488 nm, and 642 nm

CMLS-A

WAVELENGTH(S) 488 nm and 642 nm

CMLS-E

405 nm, 488 nm, 532 nm, and 642 nm

CMLS-C

488 nm, 532 nm, and 642 nm

CMLS-F

405 nm, 488 nm, 561 nm, and 642 nm

CMLS-D

488 nm, 561 nm, and 642 nm

CMLS-G

405 nm, 488 nm, 588 nm, and 642 nm

CMLS-H

488 nm and 660 nm

CMLS-I

488 nm

CMLS-J

488 nm and 532 nm

CLSD-2SS

CLSD-4SS

2-Channel Detection Module, Standard PMTs

4-Channel Detection Module, Standard PMTs

CLSD-2HS

CLSD-4HS

2-Channel Detection Module, High-Sensitivity PMTs

4-Channel Detection Module, High-Sensitivity PMTs

CLSPC-64BIT Dell 64-Bit Computer (Includes 24" Monitor)

Computer (Choose 1 Item #) CLSPC-32BIT Dell 32-Bit Computer (Includes 24" Monitor)

CLSPC-64J 64-Bit Computer with 4-Channel Digitizer (Includes 24" Monitor)

Imaging Systems Bergamo Series Microscopes

Confocal Microscopy System Choose one module from each table to configure your confocal system.

Cerna Series Microscopes

Confocal Scan Head (Choose 1) ITEM # PRICE SCAN-UV $ 35,750.00

DESCRIPTION UV/Visible Confocal Laser Scanner with Galvo-Resonant Scan Head, 16-Position Motorized Pinhole Wheel, and Two Fiber Input Ports: Port 1 Supports 488, 532, 561, and 642 nm Excitation and Port 2 Supports 405 nm Excitation

SCAN-VIS $ 33,250.00

Visible Confocal Laser Scanner with Galvo-Resonant Scan Head, 16-Position Motorized Pinhole Wheel, and One Fiber Input Port Supporting 488, 532, 561, and 642 nm Excitation

PRICE $ 12,000.00

DESCRIPTION Confocal Microscope Excitation Source for 488 nm and 642 nm

CMLS-Ba

$ 15,000.00

Confocal Microscope Excitation Source for 405 nm, 488 nm, and 642 nm

CMLS-C

$ 20,000.00

Confocal Microscope Excitation Source for 488 nm, 532 nm, and 642 nm

CMLS-D

$ 30,000.00

Confocal Microscope Excitation Source for 488 nm, 561 nm, and 642 nm

CMLS-Ea

$ 23,000.00

Confocal Microscope Excitation Source for 405 nm, 488 nm, 532 nm, and 642 nm

CMLS-Fa

$ 33,000.00

Confocal Microscope Excitation Source for 405 nm, 488 nm, 561 nm, and 642 nm

CMLS-Ga

$ 35,000.00

Confocal Microscope Excitation Source for 405 nm, 488 nm, 588 nm, and 642 nm

CMLS-H

$ 12,500.00

Confocal Microscope Excitation Source for 488 nm and 660 nm

CMLS-I

$ 11,350.00

Confocal Microscope Excitation Source for 488 nm

CMLS-J

$ 19,350.00

Confocal Microscope Excitation Source for 488 and 532 nm

aThese

Confocal Microscopes ThorImageLS Software

Laser Source (Choose 1) ITEM # CMLS-A

Essentials Kit

OCT Systems TDI Digital Microscopy Optical Tweezers

sources are only compatible with the SCAN-UV scan head.

PMT Detection Module (Choose 1) ITEM # CLSD-2SS

PRICE $ 12,250.00

DESCRIPTION 2-Channel PMT Module, Standard Sensitivity

CLSD-2HS

$ 22,750.00

2-Channel PMT Module, High Sensitivity (Includes 1 Kinematic Filter Cube Top)

CLSD-4SS

$ 21,750.00

4-Channel PMT Module, Standard Sensitivity

CLSD-4HS

$ 43,000.00

4-Channel PMT Module, High-Sensitivity (Includes 3 Kinematic Filter Cube Tops)

Data Acquisition System (Choose 1) ITEM # CLSPC-32BITa

PRICE $ 10,000.00

CLSPC-64BITb

$ 15,000.00

Dell 64-Bit Computer with 24" Monitor and 4-Channel Digitizer

CLSPC-64Jb

$ 18,000.00

High-Performance 64-Bit Computer with 24" Monitor and 4-Channel Digitizer

aThis

DESCRIPTION Dell 32-Bit Computer with 24" Monitor and 2-Channel Digitizer

computer is only compatible with systems using a 2-channel detection module. It is only compatible with ThorImageLS version1.5. computers are available with the latest version of ThorImageLS installed (version 2.0 or higher).

b64-bit

Optional Accessories The accessories below include additional PMT modules, stages for XY and Z scanning, and extra filter cubes. More information for each item is available on the pages listed in the table. ITEM # MFC1

PRICE 1,850.00

DESCRIPTION Motorized Microscope Focus Controller

PAGE 162

MLS203-1

MLS203-2

6,799.00

Fast XY Scanning Stage

168

6,799.00

Fast XY Scanning Stage for Zeiss Microscopes

MLSA01

168

155.00

MLS203 31.5 mm Riser Plates for Breadboard Mounting, 2 Pieces

MLSA02

171

105.00

Olympus IX71, IX73, and IX81 Mounting Brackets, 2 Pieces

171

MLSA03

129.00

Nikon TE2000 and Nikon Eclipse Ti Mounting Brackets, 2 Pieces

171

MLS203P2

450.00

Slide/Petri Dish Holder for Inverted Microscopes

172

BBD202

2,959.28

2-Channel Benchtop with 3-Phase Brushless DC Servo Controller

169

MZS500-E

$ 10,495.00

Z-Axis Piezo Stage and Controller Kit

175

MJC001

995.00

2-Axis Microscopy Joystick

170

MZS500P2

400.00

MZS500-Compatible Slide/Petri Dish Holder

177

MZF001

995.00

Single-Axis Joystick Controller

175

PMTSS-A

3,017.44

Amplified PMT Module

156

PMTSS2-SCM

3,200.00

Single-Channel Add-On PMT Module, Standard Sensitivity

156

PMTSS

2,300.00

Single-Channel PMT Module, Standard Sensitivity

156

PMT2000

6,750.00

High-Sensitivity GaAsP PMT Module, Cooled

158

PMT2100

6,200.00

High-Sensitivity, Compact PMT Module with Built-In Amplifier

158

DFMT1

195.00

Kinematic Dichroic Filter Cube Top

242

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes

ThorImageLS™ Software ThorImageLS Software and all required computer hardware are included with the purchase of a Thorlabs laser scanning microscopy system.

ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

ThorImageLS is a workflow-oriented image acquisition program specifically built for Thorlabs’ laser scanning microscopes. It acquires, visualizes, and analyzes images in real time; supports single image, Z-stack, time series, and image streaming acquisitions; and exports data to the lossless, metadata-preserving OME-TIFF format for compatibility with popular post-processing packages. Because it was developed under the same roof as our Bergamo II and confocal microscopes, it provides seamless and intuitive control for the entire microscopy platform.

Open-Source Software Support For users who wish to develop their own software, we provide a robust SDK that enables full access to all the functionality exposed by ThorImageLS. This SDK is employed by the ScanImage software maintained by Vidrio Technologies (formerly HHMI’s Janelia Farm), allowing customers to also use open-source packages to drive their microscopes.

Experimental Techniques n Exponential

Laser Power Ramping with Increasing Sample Depth n High-Speed Z-Stacks n Photoactivate a Selected Region of Interest (Multiphoton Dual Scan Path Systems)

Equipment Control n Position

Microscope Body and/or Sample Stages in X, Y, and Z n Average Frames and Change Pixel Dwell Times n Insert/Remove Dichroic Mirrors to Switch Between Laser Scanning and Widefield Imaging (Multiphoton Systems)

Data Analysis n Reconstruct

3D Z-Stacks n Assign a Color to Each Detection Channel n Export Data as Metadata-Tagged OME-TIFF Images or AVI Movies

Imaging Systems Bergamo Series Microscopes

New Features in ThorImageLS 2.0 ThorImageLS 2.0 incorporates significant new features to the existing ThorImageLS code base, developed in response to feedback from our users. • The Capture Setup tab (highlighted below) has been upgraded with quick-access functions that enable single-click capture of a single image, instantaneous live previews of the multiphoton image with the current microscope settings, and the ability to reset the capture settings to default. We have also added the ability to save the microscope settings as a template so that they can be recalled from disk for future acquisitions. • We have created a Script tab for automating single and multiple image acquisitions that can be immediately post processed using ImageJ macros. These powerful commands enable complex experimental sequences to be executed, tweaked, and saved for future use.

• The PMT detection channels have received their own independent panel for setting the gain, enabling frame averaging, and allowing external triggering from an electrical synchronization signal. • For Bergamo II multiphoton systems, it is now possible to save and recall up to four preset wavelengths for a Coherent Chameleon™ tunable laser.

Upgrade Eligibility For customers with supported microscopes, ThorImageLS 2.0 is available as a free upgrade. Please contact us at ImagingTechSupport@thorlabs.com to determine if your microscope is compatible with ThorImageLS 2.0. We are continuing to support ThorImageLS 1.5 for users of our older microscopes.

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Quick-Start Panel

In ThorImageLS 2.0, a panel has been added to the top of the Capture Setup tab that facilitates faster optimization of the image acquisition settings. (Image Courtesy of Dr. Hajime Hirase and Katsuya Ozawa, RIKEN Brain Science Institute, Wako, Japan)

Imaging Systems Bergamo Series Microscopes

ThorImageLS User Interface

Cerna Series Microscopes

ThorImageLS consists of a series of in-line panels that complement the hardware modules installed in the laser scanning microscope. These panels are organized into four separate tabs — Capture Setup, Capture, Review, and Script — as shown in the screenshot below. These tabs are structured to aid the user’s workflow and effectively allow the panels to be shown and hidden as needed.

Capture

Capture Setup

The Review tab is a convenient location for viewing recently acquired images. Histograms of the intensity in each detector channel as a function of pixel location are available, and sliders are provided for scrubbing along the time axis (when browsing a time series) or along the Z-axis (when examining a Z-stack). Data can be exported in the OME-TIFF format for further analysis.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

The Capture Setup tab is used to position the microscope body and sample stages, control the sample acquisition parameters, and recall previously saved settings. In multiphoton systems, it also sets the wavelength and output power (in %) of the Coherent Chameleon™ tunable laser, inserts and removes motorized dichroic mirrors, and defines the travel range and step size for Z-stack imaging. In confocal systems, this tab controls which laser sources are active, the output power (in %) of these sources, and chooses the diameter of the pinhole to be inserted into the optical path.

The Capture tab begins single and multiple image captures, including Z-stacks, time series, and photoactivation (“bleaching”) sequences. A capture can be initiated from the computer or by an external electrical synchronization signal provided by other lab equipment.

Review

Script New to ThorImageLS 2.0, the Script tab is used to program complex experimental sequences that would otherwise have to be executed manually in the other three tabs. The language consists of basic commands to capture images, run for loops, and execute ImageJ macros. More commands are planned for the future.

The right side of the window displays live images that can be zoomed and adjusted by channel intensity. 68

Imaging Systems Bergamo Series Microscopes

ThorImageLS User Interface

Cerna Series Microscopes

Selected Panels

Essentials Kit

Laser Power Control In multiphoton systems, this panel uses the Variable Attenuator and Pockels Cell (optional accessories; see pages 149 - 150 for details) to vary the laser output power, perform edge blanking, and mask out arbitrarily shaped regions. In confocal systems, this panel scales the output power of all installed excitation lasers. Laser Power Control Panel (Confocal Systems)

Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy

Laser Power Control Panel (Multiphoton Systems)

Optical Tweezers

Z Control Systems with Thorlabsâ&#x20AC;&#x2122; or Physik Instrumenteâ&#x20AC;&#x2122;s (PI) piezo objective holder installed use this panel to define the start of travel, end of travel, and number of steps in the scan for highspeed Z-stack acquisitions. This information is used to calculate a scan volume that is displayed to the user.

Z Control Panel

Tiles Control For microscopes equipped with a motorized XY stage, this panel is used to acquire several images that together span a larger area than the field of view. These images can then be stitched to generate one large image.

Tiles Control Panel

Pinhole Control (Confocal Systems Only) Our confocal microscopes use a wheel that contains 16 lithographically etched pinholes, whose circular shape prevents out-of-focus light from reaching the detectors (see page 160 for details). This panel selects the pinhole to be used and calculates, based on the objective magnification, the number of Airy Disk Units being transmitted. Pinhole Control (Confocal Systems Only) Panel

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Optical Coherence Tomography: Overview Optical Coherence Tomography (OCT) is a non-invasive, non-destructive imaging technique that enables highresolution, cross-sectional imaging of a wide range of highly scattering media, such as biological tissues. It can provide real-time 2D and 3D images (illustrated by Figures 1 and 2, respectively) of both excised samples and those inside living organisms. OCT has been employed as a valuable imaging modality in fields of scientific and medical research such as ophthalmology, angiology, neurology, dermatology, gastroenterology, embryology, oncology, and cardiology.

Telesto Series TEL113 OCT System Configuration

To address the needs for OCT systems that meet a wide range of imaging requirements, Thorlabs manufactures modular OCT components, which enable the configuration of highly customized systems. We currently offer four OCT 85.0 µm Depth product series, each of which possesses unique imaging capabilities: Vega (see pages 82 - 89), Telesto (see pages 90 - 97), Ganymede (see pages 98 - 105), and Callisto 102.0 µm Depth (see pages 106 - 113). An overview of the key parameters Figure 1. Post processing en face views of a mouse brain through the skull taken with a Telesto series OCT System. There is a separation of 17 µm for each OCT base unit is between each view. provided on page 79, followed by sample configurations of full OCT systems on pages 80 and 81. In addition, there is a configuration map at the end of each series’ presentation that outlines all the options for building a full OCT system based on that series. Figure 2. 3D volumetric image of a snail in water taken by a Telesto series OCT System.

Imaging Capabilities Thorlabs’ OCT has been used as a noninvasive imaging solution in a wide range of applications. Fields that have benefitted from our OCT technology include the following: n Small

Animal Imaging n Tissue Characterization n Live Blood Flow Imaging n Developmental Biology n Cell Imaging

n Ophthalmology n Biofilm

Inspection Imaging n Brain Activation n Non-Destructive Testing n Skin

Figure 3. Cross-sectional image of the anterior chamber of a human eye imaged with a 1050 nm Telesto series OCT system.

Advancements in Thorlabs’ OCT-based products have widely expanded OCT imaging capabilities from basic cross-sectional structural imaging to now real-time volume and micron-level imaging. Additionally, characterizing function and real-time tracking of rapid events are now common applications.

Imaging Systems Bergamo Series Microscopes

Optical Coherence Tomography: Overview

Cerna Series Microscopes

OCT Imaging Modality As an optical analog to ultrasound, OCT directs near infrared light into a sample and then measures the “time of flight” and intensity of the back reflected light. Like ultrasound, OCT non-invasively produces cross-sectional and volumetric images.

The illustration in Figure 4 to the right, outlines the advantages of three common imaging modalities used in the scientific and biomedical research fields: confocal microscopy, OCT, and ultrasound. As can be seen from the figure, OCT fills a niche between the high resolution, shallow penetration depth of confocal microscopy and the low resolution, long penetration depth of ultrasound. These unique properties of OCT can be exploited in various imaging applications.

Essentials Kit

100 µm

Resolution

Along with the non-invasive, cross-sectional imaging capability, OCT utilizes confocal gating techniques to reject backscattered light, thereby producing micron-level image resolution.

1000 µm

ras

Ult

Confocal Microscopes

d un

ThorImageLS Software

10 µm

OCT Systems

OCT

TDI Digital Microscopy

1 µm

Confocal Microscopy 0 µm 0.1 mm

Optical Tweezers 1 mm

10 mm

100 mm

Image Penetration Figure 4. OCT fills a niche between ultrasound and confocal microscopy.

Imaging a Mouse Trachea The high-speed, non-destructive, 3D imaging capability of our OCT systems provides a new approach to biomedical research. Here we demonstrate non-destructive optical sectioning of mouse trachea, ex vivo. The excised mouse trachea was maintained within a saline bath at room temperature. In this environment, the cilia that line the trachea remained active. High-speed volume images of the active trachea were acquired at 28,000 lines per second, using a Ganymede series OCT system configured with a long focal depth scan lens. As shown here, post processing of the high-resolution volume dataset allows for optical sectioning in any plane, such as en face views. Additionally, during real-time cross-sectional imaging, active cilia movement was observed. The cilia layer, goblet cell layer, and capillaries are evident in frames (b), (c), and (e), respectively.

(a)

(b)

(c)

(d)

Goblet Cell Layer

Cilia Layer

Capillaries

(e)

(f)

(g)

(h)

Figure 5. En face views of a 2 mm x 2 mm section of a mouse trachea. There is a separation of ~50 µm between each optical slice.

Imaging Systems Bergamo Series Microscopes

Optical Coherence Tomography: Overview

Cerna Series Microscopes

Cerebral Vasculature Imaging Through the Cranium

Essentials Kit

Imaging of cerebral vasculature in animal models such as mice is commonly performed to study causes for, or effects of, cerebral ischemia. Researchers at the University of Lübeck, Germany demonstrated non-invasive live mouse brain imaging through the cranium (see Figures 6 and 7 to the right) using a Telesto series OCT imaging system.

Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Figure 6. OCT en face view of a mouse cranium.

Figure 7. Angiogram OCT image of a mouse brain acquired using speckle variance imaging through the cranium.

The images in Figure 8 below show a series of en face intracranial structure and vasculature images that were also taken in this study. Each optical slice is separated by 51 μm. For this study, the high-speed, non-invasive imaging capabilities of the Telesto series system enabled acquisition of structural and functional brain images in a live mouse model. Such techniques allow long-term studies to be performed in the same animal.

153.0 µm Depth

204.0 µm Depth

255.0 µm Depth

306.0 µm Depth

357.0 µm Depth

408.0 µm Depth

459.0 µm Depth

510.0 µm Depth

561.0 µm Depth

612.0 µm Depth

Figure 8. En face intracranial vascular images overlaid on structural images of a mouse brain. Images provided by Jan Wenzel, Gereon Hüttmann, and Hendrik Spahr at the University of Lübeck.

Chick Embryo Brain Imaging

Transverse cross-sections were acquired into volumes, which enabled generation of 3D volume renderings and reconstructions, as shown to the right. Time series acquisition capabilities in Thorlabs’ OCT software provided real-time, noninvasive visualization of these changes in morphology.

Pre-Compartment Formation Post-Compartment Formation

Utilizing high-speed imaging, with long imaging depth capabilities, it is possible to track the development of organs in small animal models. Researchers at Washington University in St. Louis, Missouri employed a custom-built Thorlabs 1325 nm Swept Source OCT system to image brain formation in chicken embryo animal models before and after subdivision.

3D Renderings

3D Reconstructions

Transverse CrossSections

Figure 9. Brain formation of a chicken embryo. Reference: B.A. Filas, et al., Annals of Biomedical Engineering 39, 443-54, 2011.

Imaging Systems Bergamo Series Microscopes

Optical Coherence Tomography: Tutorial Fourier Domain Optical Coherence Tomography (FD-OCT) is based on low-coherence interferometry, which utilizes the coherent properties of a light source to measure optical path length delays in a sample. To obtain cross-sectional images with micron-level resolution using OCT, an interferometer is set up to measure optical path length differences between light reflected from the sample and reference arms. There are two types of FD-OCT systems, each characterized by its light source and detection schemes: Spectral Domain OCT (SD-OCT) and Swept Source OCT (SS-OCT). In both types of systems, light is divided by a fiber coupler into the sample and reference arms of an interferometer setup, as illustrated in Figure 10.

Cerna Series Microscopes

Spectral Domain OCT Broadband Light Source

Essentials Kit

PC FC

Reference Arm VA M

Grating

BS Sample Arm Spectrometer

CCD

G Sample

Swept Source OCT

ThorImageLS Software OCT Systems

Reference Arm

Swept Laser Source 1 3 Balanced Detector

Confocal Microscopes

CIR

C G

Sample Arm

TDI Digital Microscopy Optical Tweezers

Sample Back reflected light, attributed to variations in the index of refraction within a sample, recouples into the sample Figure 10: Schematic diagrams for the typical implementation of two Fourier Domain OCT techniques, Spectral Domain OCT and Swept Source OCT. arm fiber and then combines with the light that has FC: Fiber Coupler; PC: Polarization Controller; C: Collimator; VA: Variable traveled a fixed optical path length along the reference Attenuator; M: Mirror; CIR: Circulator; BS: Beamsplitter; G: Galvanometer arm. The resulting interferogram is measured in the detection arm of the interferometer by either a spectrometer (SD-OCT) or balanced photodetectors (SS-OCT).

The frequency of the interferogram measured by the sensor is related to the depth location of the reflector in the sample. As a result, a depth reflectivity profile (A-scan) is produced by taking a Fourier transform of the detected interferogram. 2D cross-sectional images (B-scans) are produced by scanning the OCT sample beam across the sample; by doing so, a series of A-scans are collected to create the 2D image. Similarly, when the OCT beam is scanned in a second direction, a series of 2D images is collected to produce a 3D volume dataset. With FD-OCT, 2D images are collected on time scales of milliseconds, and 3D images can be collected at rates below 1 second.

Spectral Domain OCT vs. Swept Source OCT Spectral Domain and Swept Source OCT systems are based on the same fundamental principle but incorporate different technical approaches for producing the OCT interferogram. SD-OCT systems have no moving parts and therefore have high mechanical stability and low phase noise. Availability of a broad range of line cameras has also enabled development of SD-OCT systems with varying imaging speeds and sensitivities. In contrast, SS-OCT systems utilize a wavelength swept light source and photodetector to generate rapidly the same type of interferogram. Due to the rapid sweeping of the swept laser source, high peak powers at each discrete wavelength can be used to illuminate the sample to provide greater sensitivity with little risk of optical damage.

FD-OCT Signal Processing In Fourier Domain OCT (FD-OCT), the interferogram is detected as a function of optical frequency. With a fixed optical delay in the reference arm, light reflected from different sample depths (denoted with R1, R2, and R3) produces interference patterns with different frequency components. A Fourier transform is used to resolve different depth reflections, thereby generating a depth profile of the sample (A-scan).

Figure 11. Illustration of the FD-OCT Signal Processing.

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Optical Coherence Tomography: Flow Imaging

Cerna Series Microscopes

Flow imaging is used for vessel mapping or assessing tissue function. Thorlabs provides two different methods for optical detection of flow: Doppler OCT and Speckle Variance OCT.

Essentials Kit

Doppler OCT

Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Doppler OCT is an extension of OCT that enables imaging of particle motion within a sample. This imaging capability is ideal for functional vascular imaging, studying embryonic cardiac dynamics, or monitoring vascular treatment response.

Principles of Doppler OCT In the most general terms, the Doppler effect is an apparent frequency shift of reflected waves caused by the motion of either the observer or the source, as illustrated in Figure 12 below. Although most commonly associated with sound, the Doppler effect has been observed in all types of waves, including light. A moving light source appears “bluer” (higher frequency) when approaching an observer, and “redder” (lower frequency) when moving away from one.

Figure 12. The Doppler Effect

In Doppler OCT, the Doppler effect caused by moving particles in a sample is determined by measuring a phase shift between consecutive OCT interference fringe signals. Any change in phase between consecutively acquired A-scans can be attributed to a Doppler frequency shift induced by particle motion. Thorlabs’ Doppler Imaging Mode displays the phase shift induced by moving particles using a standard Doppler color map where red to yellow indicates flow in the direction toward the OCT sample beam and violet to blue indicates flow away from the beam. This Doppler map can be clearly seen in Figure 13 below. The color map is then superimposed onto the composite B-scan to provide structural landmarks to aid in flow analysis.

Figure 13. Doppler OCT image of a rotating rod taken by a Telesto series OCT System. The red and blue color map is clearly visible on this image.

Speckle Variance OCT Speckle Variance OCT uses variation in speckle to represent particle motion. This technique is ideal for microvascular imaging because the movement of highly scattering red blood cells causes the speckle to “twinkle” like a star, allowing for easy identification. This method for optical detection of flow provides high vascular contrast while minimizing oversampling and allows visualization of flow on the cellular level. It eliminates the need for phase detection because it is sensitive to motion perpendicular to the incident beam. However, unlike Doppler OCT, Speckle Variance OCT is unable to provide information on flow orientation within the sample.

Principles of Speckle Variance OCT When a laser illuminates an irregular surface, the highly coherent light generates an interference pattern called speckle. 74

When an object moves, the speckle pattern changes, allowing for the identification of movement within a static sample. The size of the speckle is dependent on the tool being used to observe it. Thus, the size and shape of the speckle pattern does not play a role in movement identification.

Figure 14. Speckle Variance image of blood vessels taken with a Telesto series OCT System.

Speckle variance data can be overlaid on top of intensity pictures to provide morphological information. Different color maps can be used to display the multimodal pictures.

Imaging Systems

Optical Coherence Tomography: Application Support Through close collaboration with worldwide experts in OCT research and development, in conjunction with 10 years of experience in OCT design and manufacturing, Thorlabs has gained a great appreciation for the unique and often complex requirements OCT imaging applications demand. To address the individual needs of such applications, we have transformed our OCT systems into highly modular and configurable units. Modular designs, flexible configurations, and a wide variety of components and accessories allow Thorlabs’ OCT systems to be seamlessly integrated into any laboratory environment. We encourage customers to partner with our dedicated OCT application engineers to identify the optimal system configuration that meets their individual imaging requirements. Additionally, we have dedicated lab space in our Newton, NJ and Lübeck, Germany facilities where our engineers can configure an ideal OCT system and test it under specific application conditions. After testing, we provide comparative data to further guide the decision making process. For larger feasibility studies, Thorlabs also offers extensive support to validate OCT imaging processes and provides statistical data for larger sample quantities. Please contact us to discuss such potential projects.

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Thorlabs’ Headquarters in Newton, New Jersey

Optical Tweezers

Thorlabs’ Office in Lübeck, Germany

While we prefer that samples are sent directly to us so our engineers can configure an OCT system to achieve the best imaging results per customer Thorlabs’ Japan Demo Room requirements, we also welcome customers to visit any of our facilities for hands-on experience with our OCT systems. If you would like to arrange for sample testing and a technical consultation, have any questions or feedback, or need a quote, please contact us at OCT@thorlabs.com. 75

Imaging Systems Bergamo Series Microscopes

ThorImage OCT Software

Cerna Series Microscopes

ThorImage OCT is high-performance data acquisition software that is included with Thorlabs’ OCT base units and systems. This 64-bit Windows-based software package is capable of data acquisition, processing, scan control, displaying OCT images, and data archiving. Additionally, NI LabVIEW and C-based Software Development Kits (SDKs) are available.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Scan Control

Features Interactive Scan Position Control through Video Display (Draw and Scan) n A dvanced Dataset Management n H igh-Speed Volume Rendering of 3D Data n D oppler and Speckle Variance Imaging n V ersatile Scan and Acquisition Control n S oftware Development Kit n

ThorImage OCT provides numerous scan and acquisition controls. The camera integrated in the probe of our OCT system provides live video images in the application software. Defining the scan line for 2D imaging or the scan area for 3D imaging is accomplished through the easy-to-use “Draw and Scan” feature (as seen in Figure 1 below). Additionally, one can further set processing parameters, averaging parameters, and the speed and sensitivity of the device using device presets.

Dataset Management ThorImage OCT software provides advanced dataset management capabilities, which allow opening several datasets simultaneously. Datasets are uniquely defined using an identifier consisting of study (or test series) name and an experiment number. Datasets can be exported in various image formats, such as PNG, BMP, or JPG, as well as other formats suited for post-processing purposes, such as RAW/SRM, FITS, or VFF. ThorImage also features a plug-in that allows the user to open datasets in ImageJ with one click.

Off-Line Mode Figure 1. “Draw and Scan” Feature Used to Define a Scan Pattern

When an OCT Base Unit is not connected to the computer or is not powered on, ThorImage OCT will automatically load in OffLine Mode. In this mode, data files can be opened and manipulated in all the same ways as if a device was connected. Adjustment of the display parameters and exportation of datasets into different file formats, among other things, can be accomplished in this mode.

1D Mode

Figure 2. 1D Mode, A-Scan Display

In 1D mode, the OCT beam is held at one position and repeated depth scans are recorded. These depth scans can be displayed in two forms, namely A-Scan and M-Scan. The A-Scan display mode is shown in Figure 2. In this mode, the top graph shows the OCT interferogram acquired by the OCT detector, while the lower graph displays the post-processed depth/ intensity profile (A-scan).

Figure 3. 1D Mode, M-Scan Display of a Beating Snail Heart

The M-Scan display, which is shown in Figure 3, shows a series of continuously recorded A-Scans, where the y-axis represents depth and the x-axis shows the number of the A-Scans at a single position over time. The exact position of the OCT beam can be defined by a single click within the video image.

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ThorImage OCT Software 2D Mode In the 2D imaging mode, the OCT beam scans in one direction, thus acquiring cross-sectional OCT images, which are then displayed in real time (as seen in Figure 4 to the right). Line averaging before or after the Fast Fourier Transform (FFT) is available as well as B-Scan averaging. Image display parameters, such as color mapping, can be manually controlled in this mode. Automatic calculation of the optimum contrast and brightness of the displayed OCT images is also available.

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Figure 4. 2D Mode

TDI Digital Microscopy

3D Mode In the 3D imaging mode, the OCT beam sequentially scans across the sample to collect a series of 2D cross-sectional images, which are then processed to build a 3D volumetric image.

Optical Tweezers

In the ThorImage OCT software, 3D volume datasets can be viewed as orthogonal crosssectional planes and volume renderings (see Figure 5. Rendering View with Controls Shown Enlarged to the Right Figure 5). The Sectional View features crosssectional images in all three orthogonal planes, independent of the orientation in which the data was acquired. The view can be rotated as well as zoomed in and out. The Rendering View provides a volumetric rendering of the acquired volume dataset. This view enables quick 3D visualization of the sample being imaged. Planes of any orientation can be clipped to expose structures within the volume. The 3D image can be zoomed in and out as well as rotated. Furthermore, the coloring and dynamic range settings can be adjusted. Utilizing the full potential of our high-performance software in combination with our high-speed OCT base units and systems, we are able to include a Fast Volume Rendering mode in ThorImage OCT, which serves as a preview for high-resolution 3D acquisitions. In this mode, high-speed volume renderings can be displayed in real-time, providing rapid visualization of samples in three dimensions.

3D and Dataset Manager Controls Enlarged

Time Series Time Series is the newest addition to our ThorImage OCT software. This customer-inspired feature, which is shown in Figure 6, enables acquisition of an arbitrary number of 3D volume datasets over a given interval of time. Image count and imaging period are configurable through a convenient dropdown box. Each 3D image is saved in a separate file.

Time Series Controls Enlarged

Figure 6. Time Series acquisition is a new feature within the ThorImage OCT software. The controls are shown enlarged at left.

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ThorImage OCT Software

Cerna Series Microscopes

Doppler Mode

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Figure 7. Doppler OCT image of a rotating rod taken with a Telesto series OCT system.

In the Doppler mode, phase shifts between adjacent A-scans are averaged to calculate the Doppler frequency shift induced by particle motion or flow. The number of averaged lateral and axial pixels can be modified to change velocity sensitivity and resolution during phase shift calculation. The Doppler images are displayed in the main window with a color map indicating forward- or backward-directed flow, relative to the OCT beam, as depicted in Figure 7.

Speckle Variance Mode Speckle Variance Mode uses the variance of speckle noise to calculate angiographic images. It enables the visualization of three dimensional vessel trees without requiring significant blood flow or a specific acquisition speed window. The speckle variance data can be overlaid on top of intensity pictures to provide morphologic information (see Figure 8 to the right). Different color maps can be selected to display the multimodal pictures.

Figure 8. Speckle variance image of blood vessels in the back of a hand taken with a Telesto series system.

Software Development Kit Included with the ThorImage OCT user software package is a software development kit (SDK). The SDK provides a means for developing highly specialized OCT imaging software for every individual application. The SDK is available in C++ and LabVIEWTM based interfaces and includes the following:

Figure 9. Screenshot of the software development kit included with the ThorImage OCT Software.

Hardware Control

Extensive Processing Routines

Display Options

Data Import/Export Controls

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Optical Coherence Tomography: Specification Guide Center Wavelength The center wavelength of the OCT Base Unit contributes to the actual imaging depth of penetration and resolution of the system. Shorter wavelength OCT systems, such as our 830 nm, 900 nm, or 930 nm systems, are ideal for higher resolution imaging compared to our systems with center wavelengths of 1050 nm or 1300 nm. For imaging samples that have higher optical scattering properties, such as tissue, longer wavelength systems are recommended because the longer center wavelength is affected less by scattering and, therefore, the light is able to penetrate deeper into the sample and return for detection.

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A-Scan / Line Rate A single depth profile (Intensity vs. Depth) is called an A-Scan. Two-dimensional cross-sectional images, or B-Scans, are created by laterally scanning the OCT beam and collecting sequential A-scans. The speed with which a B-scan is collected depends on the A-Scan or Line Rate. For Spectral-Domain OCT systems, the A-Scan rate is determined by the frame rate of the camera in the detection spectrometer. For Swept-Source OCT systems, it is determined by the sweep repetition rate of the swept laser source.

OCT Systems TDI Digital Microscopy Optical Tweezers

Axial (Depth) Resolution In OCT, the axial resolution is proportional to the square of the center wavelength of the source divided by the source bandwidth. In practice, the axial resolution is reduced by the index of refraction (n) of the sample. For example, the axial resolution of an OCT system based on our TEL1300V2-BU Telesto Series Base Unit is <6 μm in air or <4 μm in samples such as tissue (n=1.4).

Imaging Depth Range The imaging depth range of an OCT system is determined by the design of the OCT Base Unit. This parameter identifies the maximum achievable imaging depth for the system. As with axial resolution, the actual imaging depth range is reduced by the index of refraction of the sample. Further, the actual depth of penetration will depend on the optical properties of the sample being imaged.

Striking a Balance... Thorlabs offers a wide variety of OCT systems, each one specifically designed to optimally balance its allaround performance. While we strive to push the limits in OCT capabilities, there are often tradeoffs that need to be made with regards to performance. The most common tradeoffs are among A-Scan Line Rate, Axial Resolution, and Imaging Depth Range. Below we have illustrated where each of our OCT base units lie on a scale for these three parameters. Thorlabs Base Unit

OCS1310V2-BU

OCS1310V1-BU

A-Scan Line Rate

200 kHz

100 kHz

TEL1325LV2-BU TEL1300V2-BU TEL1050V2-BU

Up to 76 kHz

GAN930V2-BU GAN905HV2-BU

Up to 36 kHz

CAL930V1-BU CAL830LSV1-BU

1.25 kHz

Thorlabs Base Unit

Axial Resolution in Tissue (n=1.4)

CAL830LSV1-BU

19 µm

OCS1310V2-BU TEL1050V2-BU OCS1310V1-BU

13 µm

TEL1325LV2-BU

8.6 µm

11.4 µm

CAL930V1-BU

5 µm

GAN930V2-BU TEL1300V2-BU

4.3 µm 3.9 µm

GAN905HV2-BU

2.1 µm

Thorlabs Base Unit

Imaging Depth Range in Air (n=1.0)

OCS1310V1-BU

12 mm

TEL1050V2-BU

7.8 mm

TEL1325LV2-BU CAL830LSV1-BU OCS1310V2-BU

7 mm 6.5 mm 5 mm

TEL1300V2-BU GAN930V2-BU

3.5 mm 2.9 mm

GAN905HV2-BU CAL930V1-BU

1.9 mm 1.7 mm 79

Imaging Systems Bergamo Series Microscopes

OCT System Example Configurations

Cerna Series Microscopes

Vega Series Configuration VEG113

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy

The Vega Series VEG113 has been configured for long depth range imaging applications such as embryology, motion tracking, and eye imaging. The Vega VEG113 system includes the following components: • 1300 nm, 100 kHz, 12 mm Depth Range Swept Source Vega Base Unit (OCS1310V1-BU) with Computer, Data Acquisition Electronics, and Pre-Installed ThorImage OCT Software • Vega Series Rigid Scanner (OCTG-1300NR) • General-Purpose Scan Lens Kit (OCT-LK3)

• Adjustable Ring Z-Spacer (OCT-AIR3) • Stand (OCT-STAND) • XY Translation and Rotation Stage (OCT-XYR1)

Alternative Vega Series Options

Optical Tweezers

n High-Speed

Vega Base Unit n Adjustable Cage-Compatible Scanner or Hand-Held Probe n Long-Depth-of-Focus or High-Resolution Scan Lens Kit n Adjustable Immersion Z-Spacer n Foot Switch

Telesto Series Configuration TEL113 The Telesto Series TEL113 has been configured for high-resolution imaging applications such as developmental biology, angiography, eye imaging, flow imaging, small animal imaging, and skin imaging. The TEL113 system includes the following components: • 1300 nm Spectral Domain Telesto Base Unit (TEL1300V2-BU) with Computer, Data Acquisition Electronics, and Pre-Installed ThorImage OCT Software • Rigid Scanner (OCTG-1300)

• General-Purpose Scan Lens Kit (OCT-LK3) • Adjustable Immersion Z-Spacer (OCT-IMM3) • Stand (OCT-STAND) • XY Translation and Rotation Stage (OCT-XYR1)

Alternative Telesto Series Options n Long

Imaging Range Telesto Base Unit n Adjustable Cage-Compatible Scanning System or Hand-Held Probe n Long-Depth-of-Focus or High-Resolution Scan Lens Kit n Adjustable Ring Z-Spacer n Foot Switch 80

Imaging Systems

OCT System Example Configurations

Cerna Series Microscopes

Ganymede Series Configuration GAN135 The Ganymede Series GAN135 has been configured for high-resolution imaging applications such as skin imaging and large-area hand-held material inspection. The Ganymede GAN135 system includes the following components: • 930 nm Ganymede Base Unit (GAN930V2-BU) with Computer, Data Acquisition Electronics, and Pre-Installed ThorImage OCT Software • Hand-Held Probe (OCTH-900)

Bergamo Series Microscopes

• General-Purpose Scan Lens Kit (OCTH-LK20-BB) • Adjustable Z-Spacer (OCTH-AIR20) • Foot Switch (OCT-PEDAL2)

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Alternative Ganymede Series Options

Optical Tweezers

n High-Resolution

Ganymede Base Unit n Rigid or Adjustable Cage-Compatible Scanner n Long-Depth-of-Focus or High-Resolution Scan Lens Kit n Adjustable Immersion Z-Spacer

Callisto Series Configuration CAL224 The Callisto Series CAL224 has been configured for optical integrators who require flexibility in adjusting and modifying the optical paths of the OCT reference and/or sample arms. The Callisto CAL224 system includes the following components: • 830 nm Long Imaging Range Callisto Base Unit (CAL830LV2-BU) with Laptop Computer and Pre-Installed ThorImage OCT Software • Adjustable Cage-Compatible Scanner (OCTP-900)

• Long-Depth-of-Focus Scan Lens Kit (OCT-LK4-BB) • Adjustable Immersion Z-Spacer (OCT-IMM4) • Stand (OCT-STAND) • Foot Switch (OCT-PEDAL2)

Alternative Callisto Series Options n Standard

930 nm Callisto Base Unit

n Rigid

Scanner or Hand-Held Probe n High-Resolution or General-Purpose Scan Lens Kit n Adjustable Ring Z-Spacer n XY Translation and Rotation Stage

Imaging Systems Bergamo Series Microscopes

Vega Series Swept Source OCT Systems

Cerna Series Microscopes

Features n Long

Imaging Range Up to 12 mm n 2D Cross-Sectional Imaging up to 200,000 Lines per Second n High-Speed 2D and 3D Acquisition n 12 µm Resolution in Biological Samples n Includes Base Unit, Data Acquisition, Computer, and Software

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Vega Series VEG113 OCT System Configuration

Thorlabs’ Vega series of configurable Swept Source Optical Coherence Tomography (OCT) systems are designed for use in applications where speed or long range imaging are the highest priority. These systems incorporate a light source that is built upon Thorlabs’ patented Micro-Electro-Mechanical (MEMS)- tunable Vertical Cavity Surface Emitting Laser (VCSEL) technology discussed on pages 196 – 199. Choose from two base units, three scanning systems, three lens kits, and additional accessories to build a tailored Vega configuration that meets your long range and high-speed OCT imaging needs.

Figure 1. En face OCT images of a plant leaf acquired with a Vega series OCT system operating at a 200 kHz line rate. Image Size: 6 mm (L) x 6 mm (W) x 100 µm Step (D).

Options at a Glance Scanning Systems • Rigid Scanner • Adjustable Cage-Compatible Scanner • Hand-Held Probe n Scan Lens Kits • Long Depth of Focus • General Purpose • High Resolution n

Figure 2. Cross-sectional OCT image of a human finger acquired using a Vega Series OCT

System operating at a 200 kHz A-Scan rate. Image Size (W x D): 15 mm x 7.4 mm (1024 A-Scans, 1024 Points per A-Scan)

Sample Z-Spacers • Adjustable Ring • Adjustable Immersion n Accessories • Scanner Stand • XY Translation and Rotation Stage • Foot Switch Activation n Functional Additions n

• Polarization-Sensitive Module 82

Imaging Systems Bergamo Series Microscopes

Vega Series Swept Source OCT Systems

OCS1310V1-BU Vega Series Base Unit

The imaging performance of any OCT system is largely dependent on the design and components incorporated into the base unit. All of Thorlabs’ OCT Base Units include an OCT “engine,” computer, software, and a software development kit (SDK). For the Vega series, the OCT “engine” is comprised of a MEMS-VCSEL light source, interferometer, scanning electronics, and a detection system. The following pages describe the various options available to configure a complete OCT system using a Vega series base unit. For a fully operational system, one scanning option and a scan lens kit must be purchased along with a base unit.

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Step #1: Choose a Base Unit While custom base unit configurations are available, we currently offer two standard Vega base unit options (described below) that have been optimized for imaging depth range and speed, respectively. Both operate at a center wavelength of 1300 nm, ideal for imaging in highly scattering and water-rich samples such as tissue.

OCS1310V1-BU

OCS1310V2-BU

Long-Range Imaging

High-Speed Imaging

The Vega series OCS1310V1-BU base unit offers the longest imaging depth range capability currently available on the market (12 mm in air). Although in most applications the actual depth of penetration will be dominated by the optical absorption and scattering properties of the sample, it is practically guaranteed that hardware limitations in imaging depth will not be a concern with this Vega base unit.

The Vega series OCS1310V2-BU base unit offers the highest speed OCT imaging capability currently available on the market, boasting 200,000 lines per second imaging. For applications such as live small animal imaging, this base unit provides the imaging speeds necessary to track dynamic processes. Additionally, the high scanning speed greatly reduces motion artifacts during 3D imaging.

VEGA SERIES OPTICAL COHERENCE TOMOGRAPHY BASE UNIT SPECIFICATIONS ITEM #

OCS1310V1-BU

Center Wavelength Sensitivity Axial/Depth Resolution (Air/Tissuea)

OCS1310V2-BU 1300 nm

>103 dB

>100 dB

16 µm / 11.4 µm

18 µm / 13 µm

Imaging Depth Range (Air)

12 mm

>5 mm

A-Scan/Line Rate

100 kHz

200 kHz

Base Unit Dimensions

Swept Laser Source: 321 mm x 320 mm x 150 mm (12.6" x 12.6" x 5.9") Imaging Module: 321 mm x 320 mm x 65 mm (12.6" x 12.6" x 2.6")

COMPUTER SPECIFICATIONSb Operating System

Windows 7, 64 Bit

Processor

Six Core

Processor Speed

2.3 GHz

Memory

32 GB DDR3 RAM

Hard Drive Data Acquisition

1 TB A/D Conversion Rate: 500 MS/s A/D Resolution: 12 Bit

aTheoretical bComputer

Value Calculated from the Refractive Index Specifications Subject to Change

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Vega Series Swept Source OCT Systems Step #2: Choose a Scanner Thorlabs’ OCT scanning systems are designed to scan the OCT light source beam across a sample in one or two dimensions, enabling 2D cross-sectional or 3D volume imaging, respectively. OCT applications can vary widely, with each requiring a different set of scanning parameters. We currently offer three scanning options for use with our Vega base units: a Rigid Scanner, an Adjustable Cage-Compatible Scanner, and a newly designed Hand-Held Probe, which is featured on page 87. All scanning systems include an integrated video camera that provides live en face video images. Our Rigid and Adjustable Cage-Compatible Scanners provide widefield, rapid scanning of the OCT beam across a sample in the X and Y directions. The Vega series-compatible Rigid Scanner (OCTG- OCTG-1300NR 1300NR) is ideal for general-purpose OCT imaging applications, as the entire design is contained within a rugged, light-tight housing that prevents risk of misalignment. For OCT integrators that require flexibility, the Adjustable Cage-Compatible Scanner [OCTP-1300NR(/M)] incorporates 8-32 (M4) taps and enables customization of the optical beam path using Thorlabs’ cage- and SM1-compatible components.

OCT Systems TDI Digital Microscopy Optical Tweezers

OCTP-1300NR

Coming Soon... Hand-Held Probe Scanning System

All scanning systems also contain a ring of LEDs around the exit aperture. This LED ring provides user-adjustable white-light illumination that aids viewing the sample through an integrated video camera.

Step #3 Choose a Scan Lens Kit

Thorlabs’ Scan Lens Kits enable easy exchange of scan lenses for flexibility in resolution, depth-of-focus, and working distance. Based on our line of OCT telecentric scan lenses (see pages 202 - 205 for information), these kits include an “LSM” scan lens, illumination tube, IR card, OCT-LK2 OCT-LK3 OCT-LK4 and calibration target. The illumination tube serves as a light guide that channels the LED ring light emanating from For a Sneak the exit aperture of the scanning system down to the sample area being imaged. Please note that the working Peak, See distance is decreased on the edge of the illumination tube compared to the center of the lens (see the table for Page 86 details). While included in the kit, use of the illumination tube is not required. The IR card and calibration target are provided to guide calibration of the scanning mirrors for use ITEM # OCT-LK2 OCT-LK3 OCT-LK4 with the associated scan lens and to ensure the best image quality when Design Wavelength 1300 nm swapping between scan lenses. Lateral Resolution* 7 µm 14 µm 20 µm Focal Length

18 mm

36 mm

54 mm

Working Distance (On Center/On Edge)

7.5 mm/ 3.3 mm

25.1 mm/ 24.8 mm

42.3 mm/ 41.6 mm

Field of View (L x W)

6 mm x 6 mm

10 mm x 10 mm

16 mm x 16 mm

*1/e2 Beam Diameter at Focus

Three Scan Lens Kits are available for use with the Vega series scanners highlighted above. Due to differences in optical paths between scan lenses, a custom sample arm patch fiber is required when interchanging scan lens kits in the Vega Series. Please contact us for more details.

Step #4: Sample Z-Spacers (Optional) Thorlabs offers both ring-like and immersion-type Sample Z-Spacers that enable optimal positioning of a scanning system relative to the sample. They can be adjusted and then locked in place for increased stability. The OCT-AIR3 ring version, which is compatible with the OCT-LK3 Scan Lens Kit, provides a distance guide between the scanner and sample without being in contact with the scanning area; only the edge is in contact with the sample.

OCT-AIR3

OCT-IMM4

Imaging Systems Bergamo Series Microscopes

Vega Series Swept Source OCT Systems We also offer immersion spacers that can be used with either our general-purpose or long-depth-offocus scan lens kits. Both consist of a glass plate that comes in contact with the sample surface within the scanning area. These immersion Z-spacers provide sample stabilization, reduce strong back reflections from the surface by allowing better refractive index matching, and enable access to samples contained within a liquid environment. OCT-IMM3 OCT-STAND

Step #5: Stand and Stage (Optional) For convenient mounting of our Rigid or Adjustable Cage-Compatible Scanners, we offer a stand that is ideal for use in vibration-sensitive studies such as angiography. It consists of a post-mounted focus block that provides both coarse and fine Z-axis travel. The focus block attaches to a specially designed 12" x 14" aluminum breadboard using an included Ø1.5" post.

Side Grips and Rubber Feet for Easy Lifting

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

OCT-XYR1 Precise translation and rotation is often required for optimal positioning of a sample before and during OCT imaging. Our OCT-XYR1 Stage provides 1" of travel in both the X and Y directions as well as 360° rotation. It also includes a solid cover plate for effortless sample mounting and easy cleaning.

Step #6: Foot Switch (Optional) Thorlabs offers a two-pedal foot switch for times when it is inconvenient to hand operate imaging software on a computer. This customer-inspired accessory, which is useful for imaging on the far end of a benchtop or when manipulating a sample, has a 2 m long USB cable and is designed to activate (and/or deactivate) two user-defined functions such as start/stop scanning and acquiring snapshots or streaming data to a disk. It seamlessly integrates with our ThorImage OCT software (see pages 76 - 78 for details).

OCT-PEDAL2

Vega Base Unit Options (Choose One) ITEM # OCS1310V1-BU

PRICE 60,000.00

DESCRIPTION Vega Series Base Unit, 100 kHz, 12 mm Imaging Depth Range

OCS1310V2-BU

63,000.00

Vega Series Base Unit, 200 kHz, 5 mm Imaging Depth Range

Scanner Options (Choose One)* ITEM # OCTG-1300NR

METRIC ITEM # –

PRICE 7,500.00

OCTP-1300NR

OCTP-1300NR/M

9,000.00

DESCRIPTION Rigid Scanner, Vega Series, 1300 nm

To Add a PolarizationSensitive Module,

Adjustable Cage-Compatible Scanner, 8-32 (M4) Taps,Vega Series, 1300 nm

*The Hand-Held Probe Presented on Page 87 can be Chosen in Lieu of One of These Options

Scan Lens Kit Options (Choose One)* ITEM # OCT-LK2

PRICE 2,000.00

DESCRIPTION High-Resolution Scan Lens Kit, LSM02

OCT-LK3

1,400.00

General-Purpose Scan Lens Kit, LSM03

OCT-LK4

1,400.00

Long-Depth-of-Focus Scan Lens Kit, LSM04

See Page 86

* If Choosing a Hand-Held Probe, Select from the Scan Kit Options Presented on Page 87

Sample Z-Spacers (Optional)* ITEM # OCT-AIR3

PRICE 700.00

DESCRIPTION Adjustable Ring Z-Spacer for OCT-LK3(-BB) General-Purpose Scan Lens Kit

OCT-IMM3

850.00

Adjustable Immersion Z-Spacer for OCT-LK3(-BB) General-Purpose Scan Lens Kit

OCT-IMM4

950.00

Adjustable Immersion Z-Spacer for OCT-LK4(-BB) Long-Depth-of-Focus Scan Lens Kit

*If Choosing a Hand-Held Probe, Choose from the Z-Spacer Options Presented on Page 87.

Stand and Stage (Optional) ITEM # OCT-STAND

PRICE 2,000.00

OCT-XYR1

700.00

DESCRIPTION Stand for Rigid and Adjustable Cage-Compatible Scanners XY Translation and Rotation Stage

Foot Switch (Optional) ITEM # OCT-PEDAL2

PRICE 200.00

DESCRIPTION USB Foot Switch, Two Pedals

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes

Vega Series Swept Source OCT Systems Step #7: Polarization-Sensitive OCT Module (Optional) PS-OCT System Features

Essentials Kit

n Add-On

Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

PSOCT-1310V1

Polarization-Sensitive Optical Coherence Tomography (PS-OCT) is a cross-sectional birefringence imaging method, ideal for a wide range of biological and engineered materials. PS-OCT is an extension of OCT that is based on measuring the polarization properties of light collected from birefringent samples. Birefringent materials decompose light into two polarization states with an optical delay being imposed on one state. Birefringence only occurs if the material is anisotropic. Materials that exhibit birefringence properties include tissues such as tendons, muscles, teeth, bones, blood vessels, and skin. In samples such as these, PS-OCT provides additional contrast compared to conventional OCT structural images as seen in the images above. The real-time, highresolution imaging capability of PS-OCT makes it well suited for studying dental abnormalities, burn depths in the skin, and material deformations.

Module for Our OCS1310V1-BU Vega Series Base Unit n Contrast Based on Birefringence of Material n Ideal for Imaging Biological Materials n Real-Time Display of Phase Retardation Images

(a)

255

0 (b)

OCT structural (a) and PS-OCT phase retardation (b) images of an oxtail sample. The strong birefringence seen in the phase retardation image indicates that highly organized structures such as collagen fibrils exist in the tissue layers.

Thorlabs has developed a real-time, fiber-based swept source PS-OCT add-on module that provides simultaneous cross-sectional imaging of the intensity and phase retardation of light backscattered from birefringent samples. This module is compatible with our Vega series OCS1310V1-BU base unit and can be incorporated at any time.

PS-OCT Images Imaging of a human fingernail bed, a chicken muscle, and a plastic component are shown below. The birefringence of the human fingernail bed and chicken muscle is due to intrinsic ordering of specific areas of biological tissue, while that of the component is induced by residual strain resulting from the manufacturing process. Note the significantly improved contrast in the phase retardation images (b) compared to the intensity-only images (a) typical of conventional OCT systems.

(a)

(b)

Fingernail

Chicken Muscle

Plastic Component

Polarization-Sensitive OCT Module (Optional)* ITEM # PSOCT-1310V1 *This

PRICE 20,000.00

Option is Only Available with the OCS1310V1-BU Vega Series Base Unit

DESCRIPTION Vega Series Polarization-Sensitive OCT Module

Imaging Systems Bergamo Series Microscopes

Vega Series Swept Source OCT Systems Hand-Held Probe

Cerna Series Microscopes

As the newest addition to the OCT scanning system options, Thorlabs will soon be releasing the OCTH-1300NR compact, light-weight hand-held probe. This customer-inspired probe provides ergonomic handling for imaging applications where mobility is required.

Essentials Kit Confocal Microscopes

Like our other scanning systems, this hand-held option includes an integrated video camera to provide real-time video imaging of samples during OCT acquisition as well as an LED illumination ring. Easy access buttons located directly on the probe enable finger-tip control of our ThorImage OCT Software (See Pages 76 - 78 for Details).

ThorImageLS Software

OCTH-1300NR Shown with the OCTH-LK20 Scan Lens Kit and the OCTH-AIR20 Z-Spacer

OCT Systems TDI Digital Microscopy

Through the ThorImage OCT Software, hand-held probe users can select from a menu of the most common imaging and acquisition software controls to “program” each of the probe buttons.

Optical Tweezers

Described below are our current hand-held probe-compatible Scan Lens Kits and Z-Spacers. We recognize that applications that benefit from a compact hand-held probe often have varying requirements in probe design. Therefore, whether it is form, fit or function, we encourage customers to discuss with our engineers their specific requirements.

Scan Lens Kits for the OCTH-1300NR Hand-Held Probe Initially, we will be offering two hand-held probe-compatible Scan Lens Kits: the OCTH-LK20 General-Purpose Scan Lens Kit and the OCTH-LK30 Long-Depth-of-Focus Scan Lens Kit. Due to the unique design of these kits, the optical path lengths are closely matched between each other, thereby eliminating the need for a matching sample arm fiber when interchanging between them.

OCTH-LK20 ITEM #

Like the Scan Lens Kits for the Rigid and Adjustable Cage-Compatible Scanners presented on page 84, these kits include an attached illumination tube, IR card, and calibration target. For the hand-held probe scan lens kit, however, the illumination tube is not removable. Details on these kits are provided in the table to the right.

OCTH-LK30 OCTH-LK20

Design Wavelength

OCTH-LK30

1300 nm

Lateral Resolution*

16.0 µm

24.0 µm

Focal Length

19.7 mm

30.2 mm

Working Distance

11.9 mm

23.4 mm

5 mm x 5 mm

8.5 mm x 8.5 mm

Field of View (L x W)

*1/e2 Beam Diameter at Focus

Sample Z-Spacers for the OCTH-1300NR Hand Held Probe A common challenge in using hand-held probes for OCT imaging is maintaining the optimal distance between the probe and the sample surface. If the distance between the probe and sample is too large, the image will fall outside of the imaging depth range of the OCT system. To ease the challenge of positioning and maintaining the hand-held OCTH-AIR20 OCTH-AIR30 probe at the optimal distance from a sample, we offer ring Z-Spacers for both the OCTH-LK20 and OCTH-LK30 Scan Lens Kits featured above. The OCTH-AIR20 and OCTH-AIR30 are specially designed to match our General-Purpose and LongDepth-of-Focus Scan Lenses, respectively. For fine tuning the distance between the probe and the sample, these ring Z-Spacers have a knurled edge which, when rotated, will adjust the working distance of the probe. Adjustable Immersion-type Z-Spacers are also under development. Please contact us for more information. 87

Vega Series OCT Systems

OCTG-1300NR Rigid Scanner

OCTH-1300NR Hand-Held Probe

Step 2:

VEG224 This Vega Series Configuration consists of a 200 kHz high-speed Swept Source base unit, adjustable cage-compatible scanner, long-depth-of-focus scan lens kit, adjustable immersion Z-spacer, and stand.

Choose a Scanning System

OCTP-1300NR(/M) Adjustable CageCompatible Scanner

Step 1:

Choose a Base Unit

Step 6:

Polarization Diversity (Optional with the OCS1310V1-BU Base Unit Only)

OCS1310V2-BU For High-Speed Imaging Applications

OCS1310V1-BU For Long-Range Imaging Applications

VEGA SERIES OCT BASE UNIT SPECIFICATIONS ITEM #

OCS1310V1-BU

Center Wavelength Sensitivity

OCS1310V2-BU

1300 nm >103 dB

>100 dB

16 µm / 11.4 µm

18 µm / 13 µm

Imaging Depth Range (Air)

12 mm

>5 mm

A-Scan/Line Rate

100 kHz

200 kHz

Base Unit Dimensions

Swept Laser Source: 321 mm x 320 mm x 150 mm Imaging Module: 321 mm x 320 mm x 65 mm

Axial/Depth Resolution (Air/Tissue)

COMPUTER SPECIFICATIONS* Operating System

Windows 7, 64 Bit

Processor

Six Core

Processor Speed

2.3 GHz

Memory Hard Drive Data Acquisition

32 GB DDR3 RAM 1 TB 500 MS/s, 12 Bit

*Computer Specifications Subject to Change PSOCT-1310V1 Polarization-Sensitive Module

The Options OCTH-LK20 General Purpose Hand-Held Probe

Scan Lens Kit

OCT-LK2 High Resolution

OCTH-LK30 Long Depth of Focus

Matching Fiber

OCT-FA2

Rigid & Adjustable Cage-Compatible Scanners OCT-LK3 General Purpose

OCT-FA3

Step 3:

Choose a Scan Lens Kit

OCT-LK4 Long Depth of Focus

Step 4:

VEG113 This Vega Series Configuration consists of a 100 kHz swept source base unit, rigid scanner, general-purpose scan lens kit, adjustable Z-spacer, stand, and stage.

OCT-FA4

Hand-Held Probe (Ring)

Choose a Z-Spacer (Optional)

OCTH-AIR20 for OCTH-LK20 Kit

OCTH-AIR30 for OCTH-LK30 Kit OCT-AIR3 for OCT-LK3 Kit (Ring)

Rigid & Adjustable Cage-Compatible Scanners

Step 5:

Choose Accessories (Optional)

OCT-XYR1 XY Translation and Rotation Stage

OCT-PEDAL2 Foot Switch

OCT-STAND Stand for Rigid and Adjustable Cage-Compatible Scanners

OCT-IMM3 for OCT-LK3 Kit (Immersion)

OCT-IMM4 for OCT-LK4 Kit (Immersion)

Imaging Systems Bergamo Series Microscopes

TelestoTM Series Spectral Domain OCT Systems

Cerna Series Microscopes

Features

Essentials Kit

n 1050

nm and 1300 nm Base Unit Center Wavelength Options n 2D Cross-Sectional Imaging Up to 76,000 Lines per Second n Four Acquisition Modes for Flexibility in Imaging Speed and Sensitivity

Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Telesto Series TEL113 OCT System Configuration

Thorlabs’ Telesto series of configurable Spectral Domain Optical Coherence Tomography (OCT) systems provides high-resolution, deep-penetration imaging. Built upon specially designed high-speed, phase-stable spectrometers that strike a balance between resolution and imaging depth, Telesto series OCT systems are ideal for high-resolution OCT

imaging of highly scattering samples, such as tissue and small animals. Choose from the three base units, three scanning systems, six lens kits, and the array of additional accessories presented on the following pages to configure a Telesto system that is tailored for use in high-resolution and phase-sensitive OCT applications.

OCT-LK3 – General Purpose Scan Lens Kit

OCT-LK2 – High-Resolution Scan Lens Kit

(b)

250 µm

Max Scan Area: 10 mm x 10 mm; Lateral Resolution in Focus: 13 µm

OCT-LK4 – Long Depth of Focus Scan Lens Kit

(a)

250 µm

Max Scan Area: 6 mm x 6 mm; Lateral Resolution in Focus: 7 µm

(c)

250 µm

Max Scan Area: 16 mm x 16 mm; Lateral Resolution in Focus: 20 µm

Figure 1. Images of a finger pad taken with a (a) high-resolution, (b) general-purpose, and (c) long-depth-of-focus Scan Lens Kit. They demonstrate the flexibility of a 1300 nm Telesto series OCT system when used with different focusing optics. Please see page 93 for details on these scan lens kits.

Options at a Glance n

Scanning Systems

Sample Z-Spacers

• Rigid Scanner

• Adjustable Ring

• Adjustable Cage-Compatible Scanner • Hand-Held Probe

• Adjustable Immersion

Scan Lens Kits • Long Depth of Focus

• Scanner Stand

• General Purpose • High Resolution 90

Accessories

• XY Translation and Rotation Stage • Foot Switch Activation

Imaging Systems Bergamo Series Microscopes

TelestoTM Series Spectral Domain OCT Systems The imaging performance of any OCT system is largely dependent on the design and components incorporated into the base unit. All of Thorlabs’ OCT Base Units include an OCT “engine,” computer, software, and a software development kit (SDK). For the Telesto series, the OCT “engine” is comprised of a superluminescent diode light source, scanning electronics, and a spectrometer-based detection system. The following pages describe the various options available to configure a complete OCT system using a Telesto series base unit. For a fully operational system, one scanning option and a scan lens kit must be purchased along with a base unit.

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems

TEL1300V2-BU Telesto Base Unit

TDI Digital Microscopy Optical Tweezers

Step #1: Choose a Base Unit While custom base unit configurations are available, we currently offer three standard Telesto base unit options (described below) that have been optimized for resolution and imaging depth range at three central wavelengths: 1050 nm, 1300 nm, or 1325 nm. These base units enable user control of imaging speed for added flexibility in sensitivity.

TEL1050V2-BU

TEL1300V2-BU

TEL1325LV2-BU

Imaging Through Water

High-Resolution Imaging

Long-Range Imaging

The Telesto series TEL1050V2-BU base unit offers imaging capabilities at a center wavelength around 1050 nm, where water absorption is significantly lower compared to 1300 nm. Using the same highsensitivity technology as in our 1300 nm Telesto series base unit, this system also offers the longest imaging depth range in this series.

The Telesto series TEL1300V2BU base unit offers Thorlabs’ highest resolution OCT imaging capability at 1300 nm. Utilizing Thorlabs’ unique matched pair of superluminescent diodes, it boasts over 170 nm of bandwidth that translates to <5.5 µm axial resolution imaging capability. The TEL1300V2-BU is the ideal choice for high-resolution imaging in scattering samples.

The Telesto series TEL1325LV2-BU base unit system offers deep imaging with good axial resolution. Like all Telesto series systems, it allows the sensitivity to be altered through adjustable A-scan rates and averaging options.

TELESTO SERIES OPTICAL COHERENCE TOMOGRAPHY BASE UNIT SPECIFICATIONS ITEM #

TEL1050V2-BU

TEL1300V2-BU

TEL1325LV2-BU

1050 nm

1300 nm

1325 nm

Center Wavelength Sensitivitya Axial/Depth Resolution (Air/Tissueb)

97 dB at Maximum A-Scan Rate 16 µm / 11.4 µm

5.5 µm / 3.9 µm

12 µm / 8.6 µm

7.8 mm

3.5 mm

7 mm

Imaging Depth Range (Air) A-Scan/Line Rate

76 kHz (at Maximum A-Scan Rate)

Base Unit Dimensions

420 mm x 320 mm x 150 mm (16.5" x 12.6" 5.9")

COMPUTER SPECIFICATIONSc Operating System

Windows 7, 64 Bit

Processor

Quad Core

Processor Speed

3.2 GHz

Memory

16 GB

Hard Drive

500 GB

Data Acquisition aUsing

a Common Path Probe with 50% Beamsplitter

Camera Link bTheoretical

Value Calculated from the Refractive Index

cComputer

Specifications are Subject to Change

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

TelestoTM Series Spectral Domain OCT Systems Step #2: Choose a Scanner Thorlabs’ OCT scanning systems are designed to scan the OCT light source beam across a sample in one or two dimensions, enabling 2D cross-sectional or 3D volume imaging, respectively. OCT applications can vary widely, from live animal imaging to analysis of excised tissue samples, with each requiring a different set of scanning parameters. We currently offer five scanning options for use with our Telesto base units: two Rigid Scanner options, two Adjustable Cage-Compatible Scanner options, and a newly designed Hand-Held Probe, which is featured on page 95. Our Rigid and Adjustable Cage-Compatible Scanners provide wide-field, rapid scanning of the OCT beam across a sample in the X and Y directions. The Telesto series-compatible Rigid Scanners are ideal for imaging applications that require a stable, easily operated setup. The entire design of the rigid scanner is contained within a rugged, lighttight housing that prevents risk of misalignment. Unlike the Vega series, Telesto series scanning systems house the reference arm path of the OCT interferometer. We have also added a reference arm path length distance indicator to this series, thereby providing easy documentation of any adjustments made to the reference arm path. Two Rigid Scanners are available: the OCTG-1300 is optimized for our 1300 nm and 1325 nm Telesto series base units (TEL1300V2-BU and TEL1325LV2-BU), while the OCTG-1050 is optimized for operation with the 1050 nm Telesto series base unit (TEL1050V2-BU). For OCT integrators that require flexibility, the Adjustable Cage-Compatible Scanning Systems [OCTP-1050(/M) and OCTP-1300(/M)] incorporate 8-32 (M4) taps and are designed for easy customization of the optical beam path using Thorlabs’ cage- and SM1-compatible components. Both the Rigid and Adjustable Cage-Compatible Scanners include a small beamsplitter and the reference arm of the OCT interferometer for easy swapping between scan lens kits. The reference arms are interchangeable. Additionally, these scanning systems contain a ring of LEDs around the exit aperture. This LED ring provides user-adjustable, white-light illumination on the sample surface that aids in visualizing the sample through the integrated video camera. When used with our ThorImage OCT software, it’s possible to obtain live en face video of the sample being imaged. Additionally, this software provides the ability to control the scan length and location from within the video image screen (see the ThorImage OCT Software presentation starting on page 76 for more details).

Coming Soon... Hand-Held Probe OCT Scanning System

OCTP-1300

For a Sneak Peak, See Page 95 92

OCTG-1050

Imaging Systems Bergamo Series Microscopes

TelestoTM Series Spectral Domain OCT Systems

Cerna Series Microscopes

Step #3 Choose a Scan Lens Kit Thorlabs’ Scan Lens Kits enable easy exchange of scan lenses for flexibility in resolution, depth-of-focus, and working distance. Based on our line of OCT telecentric scan lenses (see pages 202 - 205 for information), these kits include an “LSM” scan lens, illumination tube, IR card, and calibration target. The illumination tube serves as a light guide that channels the LED illumination ring emanating from the scanning system, down to the sample area being imaged. Please note that the working distance is decreased on the edge of the illumination tube compared to the center of the lens (see the tables for details). While included in the kit, use of the illumination tube is not required. The IR card and calibration target are provided to guide calibration of the scanning mirrors for use with the associated scan lens and to ensure the best image quality when swapping between scan lenses. Six Scan Lens Kits are available for use with the Telesto series scanners highlighted on the previous page. Three are designed for use with the 1300 nm and 1325 nm Telesto Base Units, while three others (denoted by –BB in the Item #) have a broadband coating for use with the 1050 nm Telesto Base Unit.

Essentials Kit Confocal Microscopes

OCT-LK2 ITEM #

OCT-LK3 OCT-LK2

Design Wavelength

OCT-LK4 OCT-LK3

ThorImageLS Software

OCT-LK4

1300 nm/1325 nm

Lateral Resolution*

7 µm

14 µm

20 µm

Focal Length

18 mm

36 mm

54 mm

Working Distance (On Center/On Edge)

7.5 mm/ 3.3 mm

25.1 mm/ 24.8 mm

42.3 mm/ 41.6 mm

Field of View (L x W)

6 mm x 6 mm

10 mm x 10 mm

16 mm x 16 mm

OCT-LK3-BB

OCT-LK4-BB

ITEM #

OCT-LK2-BB

Design Wavelength

OCT Systems TDI Digital Microscopy Optical Tweezers

1050 nm

Lateral Resolution*

4.5 µm

9 µm

14 µm

Focal Length

18 mm

36 mm

54 mm

Working Distance (On Center/On Edge)

7.5 mm/ 3.3 mm

25.1 mm/ 24.8 mm

42.3 mm/ 41.6 mm

Field of View (L x W)

6 mm x 6 mm

10 mm x 10 mm

16 mm x 16 mm

*1/e2 Beam Diameter at Focus

OCT-RA4

Due to the varying amount of optical material between our scan lens designs, the optical path lengths through the sample arm will also vary. Small variations, such as imaging through water, can be compensated for by adjusting the reference arm path length knob located at the top of the scanning system. When the scan lens is changed, however, a larger adjustment is required.

Step #5: Sample Z-Spacers Thorlabs offers both ring-like and immersion-type Sample Z-Spacers that enable optimal positioning of a scanning system relative to the sample. They can be adjusted and then locked in place for increased stability. The OCT-AIR3 ring version, which is compatible with the OCT-LK3 Scan Lens Kit, provides a distance guide between the scanner and sample without being in contact with the scanning area; only the edge is in contact with the sample.

OCT-IMM3

OCT-AIR3

We also offer immersion spacers, one for use with our general-purpose scan lens kit and another for use with the long-depth-of-focus kit. Both consist of a glass plate that comes in contact with the sample surface within the scanning area. These immersion Z-spacers provide sample stabilization, reduce strong back reflections from the surface by allowing better index matching, and enable access to samples contained within a liquid environment.

OCT-IMM4

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

TelestoTM Series Spectral Domain OCT Systems Step #6: Stand and Stage (Optional)

For convenient mounting of our Rigid or Adjustable Cage-Compatible Scanners presented on page 92, we offer a stand that is ideal for use in vibration-sensitive studies such as angiography. It consists of a post-mounted focus block that provides both coarse and fine Z-axis travel. The focus block attaches to a specially designed 12" x 14" aluminum breadboard using an included Ø1.5" post. The aluminum breadboard has side grips and rubber feet for easy lifting and transportation. Precise translation and rotation is often required for optimal positioning of a sample before and during OCT imaging. Our OCT-XYR1 Stage provides 1" of travel in both X and Y as well as 360° rotation. It also includes a solid cover plate for effortless sample mounting and easy cleaning. OCT-STAND OCT-XYR1

Step #7: Foot Switch (Optional) Thorlabs offers a two-pedal foot switch for times when it is inconvenient to hand operate imaging software on a computer. This customer-inspired accessory, which is useful for imaging on the far end of a benchtop or when manipulating a sample, has a 2 m long USB cable and is designed to activate (and/or deactivate) two user-defined functions. It seamlessly integrates with OCT-PEDAL2 ThorImage OCT software (see pages 76 - 78 for details). Telesto Base Units (Choose One) ITEM # TEL1050V2-BU

PRICE 63,000.00

DESCRIPTION Telesto Series Base Unit, 1050 nm

TEL1300V2-BU

59,800.00

Telesto Series Base Unit, 1300 nm

TEL1325LV2-BU

56,500.00

Telesto Series Base Unit, 1325 nm, Long Range

Scanner Options (Choose One)* ITEM # OCTG-1050

METRIC ITEM # –

PRICE 11,500.00

DESCRIPTION Rigid Scanner, 1050 nm

OCTG-1300

–

11,500.00

Rigid Scanner, 1300 nm/1325 nm

OCTP-1050

OCTP-1050/M

11,000.00

Adjustable Cage-Compatible Scanner, 8-32 (M4) Taps, 1050 nm

OCTP-1300

OCTP-1300/M

11,000.00

Adjustable Cage-Compatible Scanner, 8-32 (M4) Taps, 1300 nm/1325 nm

*The Hand-Held Probe Presented on Page 95 can be Choosen in Lieu of One of These Options

Scan Lens Kit Options (Choose One)* ITEM # OCT-LK2

PRICE 2,000.00

DESCRIPTION High-Resolution Scan Lens Kit, 1300 nm or 1325 nm, LSM02

OCT-LK3

1,400.00

General-Purpose Scan Lens Kit, 1300 nm or 1325 nm, LSM03

OCT-LK4

1,400.00

Long-Depth-of-Focus Scan Lens Kit, 1300 nm or 1325 nm, LSM04

OCT-LK2-BB

2,000.00

High-Resolution Scan Lens Kit, 1050 nm, LSM02-BB

OCT-LK3-BB

1,400.00

General-Purpose Scan Lens Kit, 1050 nm LSM03-BB

OCT-LK4-BB

1,400.00

Long-Depth-of-Focus Scan Lens Kit, 1050 nm, LSM04-BB

*If Choosing a Hand-Held Probe, Select from the Scan Kit Options Presented on Page 95

Reference Arm Adapters* ITEM # OCT-RA2

PRICE 500.00

DESCRIPTION Rigid Scanner Reference Arm Adapter for OCT-LK2(-BB) Scan Lens Kit

OCT-RA3

500.00

Rigid Scanner Reference Arm Adapter for OCT-LK3(-BB) Scan Lens Kit

OCT-RA4

500.00

Rigid Scanner Reference Arm Adapter for OCT-LK4(-BB) Scan Lens Kit

*A Reference Arm Adapter is Required with a Rigid Scanner if You Intend to Switch Scan Lenses.

Sample Z-Spacers (Optional)* ITEM # OCT-AIR3

PRICE 700.00

DESCRIPTION Adjustable Ring Z-Spacer for OCT-LK3 General-Purpose Scan Lens Kit

OCT-IMM3

850.00

Adjustable Immersion Z-Spacer for OCT-LK3 General-Purpose Scan Lens Kit

OCT-IMM4

950.00

Adjustable Immersion Z-Spacer for OCT-LK4 Long-Depth-of-Focus Scan Lens Kit

*If Choosing a Hand-Held Probe, Choose from the Z-Spacer Options Presented on Page 95

Stage and Stand (Optional) ITEM # OCT-STAND

PRICE 2,000.00

OCT-XYR1

700.00

DESCRIPTION Stand for Rigid and Adjustable Cage-Compatible Scanners XY Translation and Rotation Stage

Foot Switch (Optional) ITEM # OCT-PEDAL2

PRICE 200.00

DESCRIPTION USB Foot Switch, Two Pedals

Imaging Systems Bergamo Series Microscopes

TelestoTM Series Spectral Domain OCT Systems Hand-Held Probe

Cerna Series Microscopes

As the newest addition to the OCT scanning system options, Thorlabs will soon be releasing the OCTH-1300 compact, light-weight hand-held probe for our 1300 nm and 1325 nm Telesto Series OCT systems. This customer-inspired probe provides ergonomic handling for imaging applications where mobility is required.

Essentials Kit Confocal Microscopes

OCTH-1300 Like our other scanning systems, this hand-held option Shown with the OCTH-LK20 Scan includes an integrated video camera to provide real time Lens Kit and the OCTH-AIR20 Z-Spacer video imaging of samples during OCT acquisition and contains an LED illumination ring. Easy access buttons located directly on the probe enable finger-tip control of our ThorImage OCT Software (See Pages 76 - 78 for Details).

Through the ThorImage OCT Software, hand-held probe users can select from a menu of the most common imaging control and acquisition software controls to “program” each of the hand-held probe buttons.

ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Scan Lens Kits for the OCTH-1300 Hand-Held Probe Initially, we will be offering two hand-held probe-compatible Scan Lens Kits: the OCTH-LK20 General-Purpose Scan Lens Kit and the OCTHLK30 Long-Depth-of-Focus Scan Lens Kit. Due to the unique design of these kits, the optical path lengths are closely matched between each other, thereby eliminating the need for a separate reference arm adapter when interchanging between them. ITEM # OCTH-LK20 OCTH-LK30 Design Wavelength

1300 nm/ 1325 nm

Lateral Resolution*

16.0 µm

24.0 µm

Focal Length

19.7 mm

30.2 mm

Working Distance

11.9 mm

23.4 mm

5 mm x 5 mm

8.5 mm x 8.5 mm

Field of View (L x W)

*1/e2 Beam Diameter at Focus

OCTH-LK20

OCTH-LK30

Like the Scan Lens Kits for the Rigid and Adjustable Cage-Compatible Scanners presented on page 92, these kits include an attached illumination tube, IR card, and calibration target. For the hand-held probe scan lens kit, however, the illumination tube is not removable. Details on these kits are provided in the table to the left.

Sample Z-Spacers for OCTH-1300 Hand-Held Probe A common challenge in using hand-held probes for OCT imaging is maintaining the optimal distance between the probe and the sample surface. If the distance between the probe and sample is too large, the image will fall outside of the imaging depth range of the OCT system. OCTH-AIR20

OCTH-AIR30

To ease the challenge of positioning and maintaining the hand-held probe at the optimal distance from a sample, we offer ring Z-Spacers for both the OCTH-LK20 and OCTH-LK30 Scan Lens Kits featured above. The OCTH-AIR20 and OCTH-AIR30 are specially designed to match our General-Purpose and Long-Depth-of-Focus Scan Lenses, respectively. For fine tuning the distance between the probe and the sample, these ring Z-Spacers have a knurled edge which, when rotated, will adjust the working distance of the probe. Adjustable Immersion-type Z-Spacers are also under development. Please contact us for more information. 95

Telesto Series OCT Systems

OCTG-1300, OCTG-1050 Rigid Scanner

OCTH-1300 Hand-Held Probe

Step 2:

TEL224 The TEL224 configuration consists of a long-range imaging 1325 nm base unit, an adjustable cage-compatible scanner, long-depth-of-focus scan lens kit, and stand.

Choose a Scanning System OCTP-1050(/M), OCTP-1300(/M) Adjustable Cage-Compatible Scanner

Step 1:

Choose a Base Unit

TEL1050V2-BU For Imaging Through Water TEL1325LV2-BU For Long-Range Imaging Applications

TEL1300V2-BU For High-Resolution Imaging Applications TELESTO SERIES OCT BASE UNIT SPECIFICATIONS ITEM # Center Wavelength

TEL1050V2-BU TEL1300V2-BU TEL1325LV2-BU 1050 nm

1300 nm

Axial/Depth Resolution 16 µm / 11.4 µm 5.5 µm / 3.9 µm (Air/Tissue) Imaging Depth Range (Air)

1325 nm

97 dB at Max A-Scan Rate

Sensitivitya

7.8 mm

3.5 mm

A-Scan/Line Rate

76 kHz (at Max A-Scan Rate)

Base Unit Dimensions

420 mm x 320 mm x 150 mm

Using a Common Path Probe with 50% Beamsplitter

COMPUTER SPECIFICATIONSb

12 µm / 8.6 µm 7 mm

Operating System Windows 7, 64 Bit Processor Quad Core 3.2 GHz Processor Speed 16 GB Memory 500 GB Hard Drive Data Acquisition Camera Link b Computer Specifications Subject to Change

The Options OCTH-LK20 General Purpose

OCTH-LK30 Long Depth of Focus

Scan Lens Kits 1050 nm 1300 nm or Base Unit 1325 nm Base Unit

Hand-Held Probe

Reference Arm Adapter (Rigid Scanner Only)

Or OCT-LK2-BB High Resolution Rigid & Adjustable Cage-Compatible Scanners

OCT-LK2 High Resolution

OCT-RA2 Adapter

OCT-LK3 General Purpose

OCT-RA3 Adapter

Or OCT-LK3-BB General Purpose

Step 3:

TEL113 The TEL113 configuration consists of a high-resolution 1300 nm base unit, rigid scanner, general-purpose scan lens kit, adjustable immersion Z-Spacer, stand, and stage.

Choose a Scan Lens Kit OCT-LK4-BB Long Depth of Focus

Step 4:

OCT-LK4 Long Depth of Focus

OCT-RA4 Adapter

Hand-Held Probe (Ring)

Choose a Z-Spacer (Optional)

OCTH-AIR20 for OCTH-LK20 Kit

OCTH-AIR30 for OCTH-LK30 Kit

Rigid & Adjustable Cage-Compatible Scanner

Step 5:

OCT-AIR3 for OCT-LK3 Kit (Ring)

Choose Accessories (Optional)

OCT-XYR1 XY Translation and Rotation Stage

OCT-PEDAL2 Foot Switch

OCT-STAND Stand for Rigid and Adjustable Cage-Compatible Scanners

OCT-IMM3 for OCT-LK3 Kit (Immersion)

OCT-IMM4 for OCT-LK4 Kit (Immersion)

Imaging Systems Bergamo Series Microscopes

GanymedeTM Series Spectral Domain OCT Systems

Cerna Series Microscopes

Features n Ideal

for High-Resolution Imaging n 2 µm Axial Resolution in Tissue n 2D Cross-Sectional Imaging Up to 36,000 Lines per Second

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Ganymede Series GAN113 OCT System Configuration

Thorlabs’ Ganymede Series of configurable Spectral Domain Optical Coherence Tomography (OCT) Systems provides our highest resolution options for OCT imaging. Utilizing shorter wavelengths than the Vega and Telesto series, the Ganymede series configurations boast as low as 2 μm resolution in tissue. These systems now bridge the classical gap

between typical OCT resolutions on the order of 10 μm and the micron-level resolution capability of confocal microscopy. Choose from two base units, three scanning systems, three lens kits, and additional accessories to build a tailored Ganymede configuration that meets your high-resolution OCT imaging needs.

(a)

(b)

Figure 1. Images of a foam 3M ear plug, obtained with a Ganymede series OCT system. Frame (a) shows a 3 mm x 2 mm x 1 mm volumetric view, while Frame (b) shows a 3 mm x 2 mm x 1 mm side view slice.

Options at a Glance n

Scanning Systems

Sample Z-Spacers

• Rigid Scanner

• Adjustable Ring

• Adjustable Cage-Compatible Scanner • Hand-Held Probe

• Adjustable Immersion

Scan Lens Kits • Long Depth of Focus

• Scanner Stand

• General Purpose • High Resolution 98

Accessories

• XY Translation and Rotation Stage • Foot Switch Activation

Imaging Systems Bergamo Series Microscopes

GanymedeTM Series Spectral Domain OCT Systems

Cerna Series Microscopes

The imaging performance of any OCT system is largely dependent on the design and components incorporated into the base unit. All of Thorlabs’ OCT Base Units include an OCT “engine,” computer, software, and a software development kit (SDK). For the Ganymede series, the OCT “engine” is comprised of a superluminescent diode light source, scanning electronics, and a spectrometer-based detection system. The following pages describe the various options available to configure a complete OCT system using a Ganymede series base unit. For a fully operational system, one scanning option and a scan lens kit must be purchased along with a base unit.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy

GAN930V2-BU Ganymede Series Base Unit

Optical Tweezers

Step #1: Choose a Base Unit While custom base unit configurations are available, we currently offer two standard Ganymede base unit options (described below) that have been optimized for resolution and imaging depth range at a central wavelength of either 900 nm or 930 nm. These base units enable user control of imaging speed from 1,000 to 36,000 lines per second for added flexibility in sensitivity.

GAN930V2-BU

GAN905HV2-BU

General-Purpose Imaging

High-Resolution Imaging

The Ganymede series GAN930V2-BU base unit is designed for general-purpose OCT imaging. As a lower-cost solution over the Vega and Telesto series, the GAN930V2-BU is an ideal choice for applications that do not require the highest speeds or longest imaging depth ranges. This base unit utilizes a 930 nm superluminescent diode to provide 6 µm axial resolution in air.

The Ganymede series GAN905HV2-BU base unit provides the highest resolution OCT imaging. This spectral-domain OCT base unit utilizes a matchedpair combination of broadband superluminescent diode sources to give a 2 µm axial resolution in tissue. For applications where the highest resolution is desired, the Ganymede GAN905HV2-BU base unit is the ideal choice.

GANYMEDE SERIES OPTICAL COHERENCE TOMOGRAPHY BASE UNIT SPECIFICATIONS ITEM #

GAN930V2-BU

Center Wavelength

GAN905HV2-BU

930 nm

900 nm

Sensitivitya

91 dB (at Maximum A-Scan Rate)

Axial/Depth Resolution (Air/Tissueb)

6 µm / 4.3 µm

Imaging Depth Range (Air)

3 µm / 2.1 µm

2.9 mm

A-Scan/Line Rate

1.9 mm 36 kHz (at Maximum A-Scan Rate)

Base Unit Dimensions

420 mm x 320 mm x 150 mm (16.5" x 12.6" x 5.9")

COMPUTER SPECIFICATIONSc Operating System

Windows 7, 64 Bit

Processor

Quad Core

Processor Speed

3.2 GHz

Memory

16 GB

Hard Drive

500 GB

Data Acquisition aUsing

a Common Path Probe with 50% Beamsplitter

Camera Link bTheoretical

Value Calculated from the Refractive Index

cComputer

Specifications are Subject to Change

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

GanymedeTM Series Spectral Domain OCT Systems Step #2: Choose a Scanner Thorlabs’ OCT scanning systems are designed to scan the OCT light source beam across a sample in one or two dimensions, enabling 2D cross-sectional or 3D volume imaging, respectively. OCT applications can vary widely, from live animal imaging to analysis of excised tissue samples, with each requiring a different set of scanning parameters. We currently offer three scanning options for use with our Ganymede base units: a Rigid Scanner, an Adjustable Cage-Compatible Scanner, and a newly designed Hand-Held Probe, which is featured on page 103. Our Rigid and Adjustable Cage-Compatible Scanners provide wide-field, rapid scanning of the OCT beam across a sample in the X and Y directions. The Ganymede series-compatible Rigid Scanner (OCTG-900) is ideal for imaging applications that require a stable, easily operated setup. The entire design of the rigid scanner is contained within a rugged, light-tight housing that prevents risk of misalignment. Unlike the Vega series, Ganymede series scanning systems house the reference arm path of the OCT interferometer. We have also added a reference arm path length distance indicator to this series, thereby providing easy documentation of any adjustments made to the reference arm path. For OCT integrators that require flexibility, the Adjustable Cage-Compatible Scanner [OCTP-900(/M)] incorporates 8-32 (M4) taps and is designed for easy customization of the optical beam path using Thorlabs’ cage- and SM1compatible components. Both the Rigid and Adjustable Cage-Compatible Scanners include a small beamsplitter and the reference arm of the OCT interferometer for easy swapping between scan lens kits. The reference arms are interchangeable. Additionally, these scanning systems contain a ring of LEDs around the exit aperture. This LED ring provides user-adjustable, white-light illumination on the sample surface that aids visualizing the sample through the integrated video camera. When used with our ThorImage OCT software, it’s possible to obtain live en face video of the sample being imaged. Additionally, this software provides the ability to control the scan length and location from within the video image screen (see the ThorImage OCT Software presentation starting on page 76 for more details).

Coming Soon... Hand-Held Probe OCT Scanning System

For a Sneak Peak, See Page 103 100

OCTP-900

OCTG-900

Imaging Systems Bergamo Series Microscopes

GanymedeTM Series Spectral Domain OCT Systems Step #3 Choose a Scan Lens Kit Thorlabs’ Scan Lens Kits enable easy exchange of scan lenses for flexibility in resolution, depth-of-focus, and working distance. Based on our line of OCT telecentric scan lenses (see pages 202 - 205 for information), these kits include an “LSM” scan lens, illumination tube, IR card, and calibration target.

Cerna Series Microscopes Essentials Kit Confocal Microscopes

OCT-LK2-BB

OCT-LK3-BB

OCT-LK4-BB

The illumination tube serves as a light guide that channels the LED illumination ring emanating from the scanning system, down to the sample area being imaged. Please note that the working distance is decreased on the edge of the illumination tube compared to the center of the lens (see the table for details). While included in the kit, use of the illumination tube is not required. The IR card and calibration target are provided to guide calibration of the scanning mirrors for use with the associated scan lens and to ensure the best ITEM # OCT-LK2-BB OCT-LK3-BB Design Wavelength 900 nm/930 nm image quality when swapping between scan lenses. Three Scan Lens Kits are available for use with the Ganymede series Rigid and Adjustable Cage-Compatible Scanners: a High-Resolution Kit (OCT-LK2-BB), a General-Purpose Kit (OCT-LK3-BB), and a Long-Depth-of-Focus Kit (OCT-LK4-BB). Details are provided in the table to the right.

Lateral Resolution*

ThorImageLS Software OCT Systems TDI Digital Microscopy OCT-LK4-BB

4 µm

8 µm

12 µm

Focal Length

18 mm

36 mm

54 mm

Working Distance (On Center/On Edge)

7.5 mm/ 3.3 mm

25.1 mm/ 24.8 mm

42.3 mm/ 41.6 mm

Field of View (L x W)

6 mm x 6 mm

Optical Tweezers

10 mm x 10 mm 16 mm x 16 mm

*1/e2 Beam Diameter at Focus

Step #4: Choose a Reference Arm Adapter If you are purchasing a rigid scanner and plan to swap scan lenses, you will need to purchase the appropriate reference arm adapter. For each available scan lens kit, an associated reference arm adapter is available. In the Ganymede series, the reference arm of the OCT OCT-RA2 OCT-RA3 OCT-RA4 interferometer is housed within the scanning system, near the sample. A key aspect in the design of OCT systems is to optically match the reference and sample arm path lengths, as close as possible. This reduces any image distortion caused by dispersion. Due to the varying amount of optical material between our scan lens designs, the optical path lengths through the sample arm will also vary. Small variations, such as imaging through water, can be compensated for by adjusting the reference arm path length knob located at the top of the scanning system. When the scan lens is changed, however, a larger adjustment is required.

Step #5: Sample Z-Spacers Thorlabs offers both ring-like and immersion-type Sample Z-Spacers that enable optimal positioning of a scanning system relative to the sample. They can be adjusted and then locked in place for increased stability. The OCT-AIR3 ring version, which is compatible with the OCT-LK3-BB Scan Lens Kit, provides a distance guide between the scanner and sample without being in contact with the scanning area; only the edge is in contact with the sample.

OCT-AIR3

OCT-IMM3

OCT-IMM4

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes

GanymedeTM Series Spectral Domain OCT Systems Step #6: Stand and Stage (Optional) For convenient mounting of our Rigid or Adjustable Cage-Compatible Scanners presented on page 100, we offer a stand that is ideal for use in vibration-sensitive studies such as angiography. It consists of a postmounted focus block that provides both coarse and fine Z-axis travel. The focus block attaches to a specially designed 12" x 14" aluminum breadboard using an included Ø1.5" post. The aluminum breadboard has side grips and rubber feet for easy lifting and transportation.

ThorImageLS Software

Precise translation and rotation is often required for optimal positioning of a sample OCT-XYR1 before and during OCT imaging. Our OCT-XYR1 Stage provides 1" of travel in both the X and Y directions as well as 360° rotation. It also includes a solid cover plate for effortless sample mounting and easy cleaning.

OCT Systems TDI Digital Microscopy

OCT-STAND

Optical Tweezers

Step #7: Foot Switch (Optional)

OCT-PEDAL2

Thorlabs offers this two-pedal foot switch for times when it is inconvenient to hand operate imaging software on a computer. This customer-inspired accessory, which is useful for imaging on the far end of a benchtop or when manipulating a sample, has a 2 m long USB cable and is designed to activate (and/or deactivate) two user-defined functions. It seamlessly integrates with ThorImage OCT software (see pages 76 - 78 for details).

Ganymede Base Units (Choose One) ITEM # GAN930V2-BU

PRICE 38,000.00

GAN905HV2-BU

43,500.00

DESCRIPTION Ganymede Series Base Unit, 930 nm Ganymede Series Base Unit, 900 nm, High Resolution

Scanner Options (Choose One)* ITEM # OCTG-900

METRIC ITEM # –

PRICE 11,500.00

DESCRIPTION Rigid Scanner, 830 nm/900 nm/930 nm

OCTP-900

OCTP-900/M

11,000.00

Adjustable Cage-Compatible Scanner, 8-32 (M4) Taps, 830 nm/900 nm/930 nm

*The Hand-Held Probe Presented on Page 103 can be Choosen in Lieu of One of These Options

Scan Lens Kit Options (Choose One)* ITEM # OCT-LK2-BB

PRICE 2,000.00

DESCRIPTION High-Resolution Scan Lens Kit, LSM02-BB

OCT-LK3-BB

1,400.00

General-Purpose Scan Lens Kit, LSM03-BB

OCT-LK4-BB

1,400.00

Depth-of-Focus Scan Lens Kit, LSM04-BB

*If Choosing a Hand-Held Probe, Select from the Scan Kit Options Presented on Page 103

Reference Arm Adapters* ITEM # OCT-RA2

PRICE 500.00

DESCRIPTION Rigid Scanner Reference Arm Adapter for OCT-LK2-BB Scan Lens Kit

OCT-RA3

500.00

Rigid Scanner Reference Arm Adapter for OCT-LK3-BB Scan Lens Kit

OCT-RA4

500.00

Rigid Scanner Reference Arm Adapter for OCT-LK4-BB Scan Lens Kit

*A Reference arm Adapter is Required with a Rigid Scanner if You Intend to Switch Scan Lenses

Sample Z-Spacers (Optional)*

ITEM # OCT-AIR3

PRICE 700.00

DESCRIPTION Adjustable Ring Z-Spacer for OCT-LK3-BB General-Purpose Scan Lens Kit

OCT-IMM3

850.00

Adjustable Immersion Z-Spacer for OCT-LK3-BB General-Purpose Scan Lens Kit

OCT-IMM4

950.00

Adjustable Immersion Z-Spacer for OCT-LK4-BB Long-Depth-of-Focus Scan Lens Kit

*If Choosing a Hand-Held Probe, Choose from the Z-Spacer Options Presented on Page 103

Stand and Stage (Optional) ITEM # OCT-STAND

PRICE 2,000.00

OCT-XYR1

700.00

DESCRIPTION Stand for Rigid and Adjustable Cage-Compatible Scanners Translation and Rotation Sample Stage

Foot Switch (Optional) ITEM # OCT-PEDAL2

102

PRICE 200.00

DESCRIPTION USB Foot Switch, Two Pedals

Imaging Systems

GanymedeTM Series Spectral Domain OCT Systems

Bergamo Series Microscopes

Hand-Held Probe

Cerna Series Microscopes

As the newest addition to the OCT scanning system options, Thorlabs will soon be releasing the OCTH-900 compact, light-weight hand-held probe. This customer-inspired probe provides ergonomic handling for imaging applications where mobility is required. Like our other scanning systems, this hand-held option includes an integrated video camera to provide real time video imaging of samples during OCT acquisition and contains an LED illumination ring. Easy access buttons located directly on the probe enable finger-tip control of our ThorImage OCT Software (See Pages 76 - 78 for Details).

Essentials Kit Confocal Microscopes

OCTH-900 Shown with the OCTH-LK20-BB Scan Lens Kit and the OCTH-AIR20-BB Z-Spacer

ThorImageLS Software OCT Systems TDI Digital Microscopy

Through the ThorImage OCT Software, hand-held probe users can select from a menu of the most common imaging control and acquisition software controls to “program” each of the probe buttons.

Optical Tweezers

Scan Lens Kits for the OCTH-900 Hand-Held Probe Initially, we will be offering two hand-held probe-compatible Scan Lens Kits: a General-Purpose Kit (OCTH-LK20-BB) and a Long-Depthof-Focus Kit (OCTH-LK30-BB). Due to the unique design of these kits, the optical path lengths are closely matched between each other, OCTH-LK20-BB thereby eliminating the need for a separate reference arm adapter when interchanging between them. ITEM #

OCTH-LK20-BB

Design Wavelength Lateral Resolution* Focal Length Working Distance Field of View (L x W)

OCTH-LK30-BB

900 nm/930 nm 9.0 µm

14.0 µm

19.6 mm

30.0 mm

11.9 mm

23.4 mm

5.0 mm x 5.0 mm

8.5 mm x 8.5 mm

*1/e2 Beam Diameter at Focus

OCTH-LK30-BB

Like the Scan Lens Kits for the Rigid and Adjustable Cage-Compatible Scanners presented on page 100, these kits include an attached illumination tube, IR card, and calibration target. For the hand-held probe scan lens kit, however, the illumination tube is not removable. Details on these kits are provided in the table to the left.

Sample Z-Spacers for the OCTH-900 Hand-Held Probe A common challenge in using hand-held probes for OCT imaging is maintaining the optimal distance between the probe and the sample surface. If the distance between the probe and sample is too large, the image will fall outside of the imaging depth range of the OCT system. To ease the challenge of positioning and maintaining the hand-held OCTH-AIR20 OCTH-AIR30 probe at the optimal distance from a sample, we offer ring Z-Spacers for both the OCTH-LK20 and OCTH-LK30 Scan Lens Kits featured above. The OCTH-AIR20 and OCTH-AIR30 are specially designed to match our General-Purpose and Long-Depth-of-Focus Scan Lenses, respectively. For fine tuning the distance between the probe and the sample, these ring Z-Spacers have a knurled edge which, when rotated, will adjust the working distance of the probe. Adjustable Immersion-type Z-Spacers are also under development. Please contact us for more information. 103

Ganymede Series OCT Systems

OCTG-900 Rigid Scanner

OCTH-900 Hand-Held Probe

Step 2:

GAN135 The GAN135 configuration consists of a 930 nm base unit, hand-held probe scanning system, general-purpose scan lens kit, ring Z-spacer, and foot switch.

Choose a Scanning System OCTP-900, OCTP-900/M Adjustable Cage-Compatible Scanner

Step 1:

Choose a Base Unit

GAN930V2-BU Video-Rate, General-Purpose Imaging

GAN905HV2-BU Highest Resolution Imaging

GANYMEDE SERIES OCT BASE UNIT SPECIFICATIONS ITEM #

GAN930V2-BU

GAN905HV2-BU

Center Wavelength

930 nm

900 nm

Sensitivitya

91 dB at Maximum A-Scan Rate

Axial/Depth Resolution (Air/Tissue) Imaging Depth Range (Air) A-Scan/Line Rate a

6 µm / 4.3 µm 2.9 mm

3 µm / 2.1 µm 1.9 mm

36 kHz (at Maximum A-Scan Rate)

Base Unit Dimensions 420 mm x 320 mm x 150 mm Using a Common Path Probe with 50% Beamsplitter

COMPUTER SPECIFICATIONSb Operating System

Windows 7, 64 Bit

Processor

Quad Core

Processor Speed

3.2 GHz

Memory Hard Drive Data Acquisition

16 GB 500 GB Camera Link

Computer Specifications Subject to Change

The Options OCTH-LK20-BB OCTH-LK30-BB General Purpose Long Depth of Focus Hand-Held Probe

Scan Lens Kit

Reference Arm Adapter (Rigid Scanner Only)

OCT-LK2-BB High Resolution

OCT-RA2 Adapter

Rigid & Adjustable Cage-Compatible Scanners OCT-LK3-BB General Purpose

OCT-RA3 Adapter

Step 3:

Choose a Scan Lens Kit

OCT-LK4-BB Long Depth of Focus

Step 4:

GAN212 The GAN212 configuration consists of a high-resolution 900 nm base unit, rigid scanner, high-resolution scan lens kit, stand, and stage.

OCT-RA4 Adapter

Hand-Held Probe (Ring)

Choose a Z-Spacer (Optional)

OCTH-AIR20 for OCTH-LK20-BB Kit

OCTH-AIR30 for OCTH-LK30-BB Kit OCT-AIR3 for OCT-LK3-BB Kit (Ring)

Rigid & Adjustable Cage-Compatible Scanners

Step 5:

Choose Accessories (Optional)

OCT-XYR1 XY Translation and Rotation Stage

OCT-PEDAL2 Foot Switch

OCT-STAND Stand for Rigid and Adjustable Cage-Compatible Scanners

OCT-IMM3 for OCT-LK3-BB Kit (Immersion)

OCT-IMM4 for OCT-LK4-BB Kit (Immersion)

Imaging Systems Bergamo Series Microscopes

CallistoTM Series Spectral Domain OCT Systems

Cerna Series Microscopes

Features

Essentials Kit

n High-Sensitivity

for Increased Contrast OCT Imaging n Ideal for Imaging Static or In Vitro Samples n Low-Cost and Portable OCT Imaging

Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Callisto Series CAL113 OCT System Configuration

Thorlabs’ Callisto Series of configurable Spectral Domain Optical Coherence Tomography (OCT) Systems offer superior sensitivity for increased contrast over other OCT imaging systems (refer to Figure 1 below). They are a great tool for imaging either static, excised samples or materials. They are also commonly used as an educational tool. The high-sensitivity capabilities of the Callisto series enables distinction between layers of similar material that often (a) cannot be visualized by use of other OCT systems. Operation is performed via a laptop computer, making them easily portable to move between benchtops or classrooms. (b) Choose from two base units, three scanning systems, three lens kits, and additional accessories to build a tailored Callisto configuration that meets your high-sensitivity OCT Figure 1. Comparison of a grape skin image obtained using (a) a high-sensitivity Callisto series system and (b) a high-speed but lower-sensitivity system. Image Size: 6.3 mm x 1.7 mm imaging needs.

Options at a Glance n

Scanning Systems • Rigid Scanner

• Adjustable Cage-Compatible Scanner • Hand-Held Probe

• Adjustable Immersion

Scan Lens Kits

• Long Depth of Focus • General Purpose • High Resolution 106

Sample Z-Spacers • Adjustable Ring Accessories • Scanner Stand n

• XY Translation and Rotation Stage • Foot Switch Activation

Imaging Systems Bergamo Series Microscopes

CallistoTM Series Spectral Domain OCT Systems The imaging performance of any OCT system is largely dependent on the design and components incorporated into the base unit. All of Thorlabs’ OCT Base Units include an OCT “engine,” a computer, software, and a software development kit (SDK). For the Callisto series, the OCT “engine” is comprised of a superluminescent diode light CAL930V1-BU source, scanning electronics, and a spectrometerCallisto Series Base Unit based detection system. The following pages describe the various options available to configure a complete OCT system using a Callisto series base unit. For a fully operational system, one scanning option and a scan lens kit must be purchased along with a base unit.

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Step #1: Choose a Base Unit While custom base unit configurations are available, we currently offer two standard Callisto base unit options (described below) that have been optimized for static sample imaging.

CAL930V1-BU

CAL830LSV1-BU

Low-Cost Imaging

Long Range, High-Sensitivity

The Callisto series CAL930V1-BU is our “entrylevel” OCT base unit with center wavelength of 930 nm. This base unit provides high-sensitivity OCT imaging at a considerably slow A-scan rate of 1,200 lines per second. Intended for imaging of static samples or use as an educational tool, the CAL930V1-BU will provide as low as 7 µm resolution in air. This base unit is ideal for in vitro imaging applications.

The Callisto series CAL830LSV1-BU base unit is designed to offer the high-sensitivity capabilities of the CAL930V1-BU but with an extended imaging depth range capability. With the CAL830LSV1BU, up to 6.5 mm imaging depth can be achieved, making it ideal for long-range imaging of semitransparent or low-scattering samples.

CALLISTO SERIES OPTICAL COHERENCE TOMOGRAPHY BASE UNIT SPECIFICATIONS ITEM #

CAL930V1-BU

CAL830V1-BU

Center Wavelength

930 nm

830 nm

Sensitivitya

105 dB

109 dB

7 µm / 5 µm

27 µm / 19 µm

Axial/Depth Resolution

(Air/Tissueb)

Imaging Depth Range (Air) A-Scan/Line Rate Base Unit Dimensions

1.7 mm

6.5 mm 1.25 kHz 420 mm x 320 mm x 150 mm (16.5" x 12.6" x 5.9")

COMPUTER SPECIFICATIONSc Operating System

Windows 7, 64 Bit

Processor

Dual Core

Processor Speed

2.5 GHz

Memory Hard Drive Data Acquisition

4 GB 250 GB USB

aUsing

a Common Path Probe with 50% Beamsplitter Value Calculated from the Refractive Index cComputer Specifications are Subject to Change bTheoretical

107

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

CallistoTM Series Spectral Domain OCT Systems Step #2: Choose a Scanner Thorlabs’ OCT scanning systems are designed to scan the OCT light source beam across a sample in one or two dimensions, enabling 2D cross-sectional or 3D volume imaging, respectively. OCT applications can vary widely, from live animal imaging to analysis of excised tissue samples, with each requiring a different set of scanning parameters. We currently offer three scanning options for use with our Callisto base units: a Rigid Scanner, an Adjustable Cage-Compatible Scanner, and a newly designed Hand-Held Probe, which is featured on page 111. Our Rigid and Adjustable Cage-Compatible Scanners provide wide-field, rapid scanning of the OCT beam across a sample in the X and Y directions. The Callisto series-compatible Rigid Scanner (OCTG-900) is ideal for imaging applications that require a stable, easily operated setup. The entire design of the rigid scanner is contained within a rugged, light-tight housing that prevents risk of misalignment. Unlike the Vega series, Callisto series scanning systems house the reference arm path of the OCT interferometer. We have also added a reference arm path length distance indicator to this series, thereby providing easy documentation of any adjustments made to the reference arm path. For OCT integrators that require flexibility, the Adjustable Cage-Compatible Scanner [OCTP-900(/M)] incorporates 8-32 (M4) taps and is designed for easy customization of the optical beam path using Thorlabs’ cage- and SM1-compatible components.

OCTG-900

Both the Rigid and Adjustable Cage-Compatible Scanners include a small beamsplitter and the reference arm of the OCT interferometer for easy swapping between scan lens kits. The reference arms are interchangeable. Additionally, these scanning systems contain a ring of LEDs around the exit aperture. This LED ring provides user-adjustable, white-light illumination on the sample surface that aids in visualizing the sample through the integrated video camera. When used with our ThorImage OCT software, it’s possible to obtain live en face video of the sample being imaged. Additionally, this software provides the ability to control the scan length and location from within the video image screen (see the ThorImage OCT Software presentation starting on page 76 for more details).

Coming Soon... Hand-Held Probe OCT Scanning System

OCTP-900

For a Sneak Peak, See Page 111 108

Imaging Systems Bergamo Series Microscopes

CallistoTM Series Spectral Domain OCT Systems Step #3 Choose a Scan Lens Kit Thorlabs’ Scan Lens Kits enable easy exchange of scan lenses for flexibility in resolution, depth-of-focus, and working distance. Based on our line of OCT telecentric scan lenses (see pages 202 - 205 for information), these kits include an “LSM” scan lens, illumination tube, IR card, and calibration target.

Cerna Series Microscopes Essentials Kit Confocal Microscopes

OCT-LK2-BB

The illumination tube serves as a light guide that channels the LED illumination ring emanating from the scanning system, down to the sample area being imaged. Please note that the working distance is decreased on the edge of the illumination tube compared to the center of the lens (see the table for details). While included in the kit, use of the illumination tube is not required. The IR card and calibration target are provided to guide calibration of the scanning mirrors for use with the associated scan lens and to ensure the best image quality when swapping between scan lenses. Three Scan Lens Kits are available for use with the Ganymede series Rigid and Adjustable Cage-Compatible Scanners: a High-Resolution Kit (OCT-LK2-BB), a General-Purpose Kit (OCTLK3-BB), and a Long-Depth-of-Focus Kit (OCT-LK4-BB). Details are provided in the table to the right.

OCT-LK3-BB

ITEM #

OCT-LK2-BB

OCT-LK4-BB OCT-LK3-BB

Design Wavelength

OCT-LK4-BB

ThorImageLS Software

830 nm

OCT Systems

Lateral Resolution*

3.5 µm

7 µm

11 µm

Focal Length

18 mm

36 mm

54 mm

Working Distance (On Center/On Edge)

7.5 mm/ 3.3 mm

25.1 mm/ 24.8 mm

42.3 mm/ 41.6 mm

Field of View (L x W)

6 mm x 6 mm

ITEM #

OCT-LK2-BB

TDI Digital Microscopy Optical Tweezers

10 mm x 10 mm 16 mm x 16 mm OCT-LK3-BB

Design Wavelength

OCT-LK4-BB

930 nm

Lateral Resolution*

4 µm

8 µm

12 µm

18 mm

36 mm

54 mm

Working Distance (On Center/On Edge)

7.5 mm/ 3.3 mm

25.1 mm/ 24.8 mm

42.3 mm/ 41.6 mm

Field of View (L x W)

6 mm x 6 mm

Focal Length

10 mm x 10 mm 16 mm x 16 mm

*1/e2 Beam Diameter at Focus

OCT-RA4

Step #5: Sample Z-Spacers (Optional) Thorlab offers both ring-like and immersion-type Sample Z-Spacers that enable optimal positioning of a scanning system relative to the sample. They can be adjusted and then locked in place for increased stability. The OCT-AIR3 ring version, which is compatible with the OCT-LK3-BB Scan Lens Kit, provides a distance guide between the scanner and sample without being in contact with the scanning area; only the edge is in contact with the sample.

OCT-AIR3

OCT-IMM3

OCT-IMM4

We also offer an immersion spacers, one for use with our general-purpose scan lens kit and another for use with the long-depth-of-focus kit. Both consist of a glass plate that comes in contact with the sample surface within the scanning area. These Immersion Z-spacers provide sample stabilization, reduce strong back reflections from the surface by allowing better index matching, and enable access to samples contained within a liquid environment. 109

Imaging Systems Bergamo Series Microscopes

CallistoTM Series Spectral Domain OCT Systems Step #6: Stand and Stage (Optional)

Cerna Series Microscopes

For convenient mounting of our Rigid or Adjustable Cage-Compatible Scanners presented on page 108, we offer a stand that is ideal for use in vibration-sensitive studies such as angiography. It consists of a post-mounted focus block that provides both coarse and fine Z-axis travel. The focus block attaches to a specially designed 12" x 14" aluminum breadboard using an included Ø1.5" post. The aluminum breadboard has side grips and rubber feet for easy lifting and transportation.

Essentials Kit Confocal Microscopes ThorImageLS Software

Fine translation and rotation is often required for optimal positioning of a sample before and during OCT imaging. Our OCT-XYR1 Stage provides 1" of travel in both the X and OCT-XYR1 Y directions as well as 360° rotation. It also includes a solid cover plate for effortless sample mounting and easy cleaning.

OCT Systems TDI Digital Microscopy Optical Tweezers

OCT-STAND

Step #7: Foot Switch (Optional) Thorlabs offers a two-pedal foot switch for times when it is inconvenient to hand operate imaging software on a computer. This customer-inspired accessory, which is useful for imaging on the far end of a benchtop or when manipulating a sample, has a 2 m long USB cable and is designed to activate (and/or deactivate) two user-defined functions. It seamlessly integrates with ThorImage OCT software (see pages 76 - 78 for details). OCT-PEDAL2 Callisto Base Units (Choose One) ITEM # CAL930V1-BU

PRICE 21,300.00

CAL830LSV1-BU

27,100.00

DESCRIPTION Callisto Series Base Unit, 930 nm Callisto Series Base Unit, 830 nm, Long Range, High Sensitivity

Scanner Options (Choose One)* ITEM # OCTG-900

METRIC ITEM # –

PRICE 11,500.00

OCTP-900

OCTP-900/M

11,000.00

DESCRIPTION Rigid Scanner, 830 nm / 900 nm / 930 nm Adjustable Cage-Compatible Scanner, 8-32 (M4) Taps, 830 nm / 900 nm / 930 nm

*The Hand-Held Probe Presented on Page 111 can be Choosen in Lieu of One of These Options

Scan Lens Kit Options (Choose One)* ITEM # OCT-LK2-BB

PRICE 2,000.00

DESCRIPTION High-Resolution Scan Lens Kit, LSM02-BB

OCT-LK3-BB

1,400.00

General-Purpose Scan Lens Kit, LSM03-BB

OCT-LK4-BB

1,400.00

Depth-of-Focus Scan Lens Kit, LSM04-BB

*If Choosing a Hand-Held Probe, Select from the Scan Kit Options Presented on Page 111

Reference Arm Adapters* ITEM # OCT-RA2

PRICE 500.00

DESCRIPTION Rigid Scanner Reference Arm Adapter for OCT-LK2-BB Scan Lens Kit

OCT-RA3

500.00

Rigid Scanner Reference Arm Adapter for OCT-LK3-BB Scan Lens Kit

OCT-RA4

500.00

Rigid Scanner Reference Arm Adapter for OCT-LK4-BB Scan Lens Kit

*A Reference Arm Adapter is Required with a Rigid Scanner if You Intend to Switch Scan Lenses

Sample Z-Spacers (Optional)* ITEM # OCT-AIR3

PRICE 700.00

DESCRIPTION Adjustable Ring Z-Spacer for OCT-LK3-BB General-Purpose Scan Lens Kit

OCT-IMM3

850.00

Adjustable Immersion Z-Spacer for OCT-LK3-BB General-Purpose Scan Lens Kit

OCT-IMM4

950.00

Adjustable Immersion Z-Spacer for OCT-LK4 -BB Long-Depth-of-Focus Scan Lens Kit

*If Choosing a Hand-Held Probe, Choose from the Z-Spacer Options Presented on Page 111. Note that a Z-Spacer is Required for Callisto Systems using a Hand-Held Probe.

Stand and Stage (Optional) ITEM # OCT-STAND

PRICE 2,000.00

OCT-XYR1

700.00

DESCRIPTION Stand for Rigid and Adjustable Cage-Compatible Scanners XY Translation and Rotation Stage

Foot Switch (Optional) ITEM # OCT-PEDAL2

110

PRICE 200.00

DESCRIPTION USB Foot Switch, Two Pedals

Imaging Systems Bergamo Series Microscopes

CallistoTM Series Spectral Domain OCT Systems Hand-Held Probe

Cerna Series Microscopes

Essentials Kit Confocal Microscopes

OCTH-900 Shown with the OCTH-LR20-BB Scan Lens Kit and the OCTH-AIR20 Z-Spacer

ThorImageLS Software OCT Systems TDI Digital Microscopy

Through the ThorImage OCT Software, hand-held probe users can select from a menu of the most common imaging control and acquisition software controls to “program” each of the probe buttons.

Optical Tweezers

Described below are our current hand-held probe-compatible Scan Lens Kits and Z-Spacers. Due to the slower scan rate of the Callisto series, sample Z-Spacers are required to achieve good quality OCT images with a hand-held probe. We recognize that applications that benefit from a compact hand-held probe often have varying requirements in probe design. Therefore, whether it is form, fit or function, we encourage customers to discuss with our engineers their specific requirements.

Scan Lens Kits for the OCTH-900 Hand-Held Probe Initially, we will be offering two hand-held probe-compatible Scan Lens Kits: a General-Purpose Kit (OCTH-LK20-BB) and a Long-Depth-ofFocus Kit (OCTH-LK30-BB). Due to the unique design of these kits, the optical path lengths are closely matched between each other, thereby OCTH-LK20-BB OCTH-LK30-BB eliminating the need for ITEM # OCTH-LK20-BB OCTH-LK30-BB a separate reference arm adapter when interchanging between Design Wavelength 900 nm/930 nm them. Lateral Resolution*

9.0 µm

14.0 µm

Focal Length

19.6 mm

30.0 mm

Working Distance

11.9 mm

23.4 mm

5.0 mm x 5.0 mm

8.5 mm x 8.5 mm

Field of View (L x W)

*1/e2 Beam Diameter at Focus

Like the Scan Lens Kits for the Rigid and Adjustable CageCompatible Scanners, these kits include an attached illumination tube, IR card, and calibration target. For the hand-held probe scan lens kits, however, the illumination tube is not removable. Details on these kits are provided in the table to the left.

Sample Z-Spacers for the OCTH-900 Hand-Held Probe Due to the slower scan rates, a challenge in using hand-held probes for OCT imaging with the Callisto series base unit is maintaining the optimal distance between the probe and the sample with the least amount of motion. Motion during OCT image acquisition will result in artifacts appearing in the OCT images.

OCTH-AIR20

OCTH-AIR30

To ease the challenge of positioning and maintaining the hand-held probe at the optimal distance from a sample, we offer ring Z-Spacers for both the OCTH-LK20-BB and OCTH-LK30-BB Scan Lens Kits. The OCTH-AIR20 and OCTH-AIR30 are specially designed to match our General-Purpose and Long-Depthof-Focus Scan Lenses, respectively. For fine tuning the distance between the probe and sample, these ring Z-Spacers have a knurled edge which, when rotated, will adjust the working distance of the probe. Immersiontype Z-Spacers are also under development. Please contact us for more information. 111

Callisto Series OCT Systems

OCTG-900 Rigid Scanner

OCTH-900 Hand-Held Probe

Step 2:

CAL123 The CAL123 configuration consists of a 930 nm base unit, adjustable cage-compatible scanner, general-purpose scan lens kit, and stand.

Choose a Scanning System OCTP-900(/M) Adjustable Cage-Compatible Scanner

Step 1:

Choose a Base Unit

CAL930V1-BU Low Cost Imaging Ideal for Static Samples or Education

CAL830LSV1-BU Long-Range, High-Sensitivity Imaging of Static Samples

CALLISTO SERIES OCT BASE UNIT SPECIFICATIONS ITEM #

CAL930V1-BU

CAL830LSV1-BU

Center Wavelength

930 nm

830 nm

Operating System

Sensitivitya

105 dB

109 dB

Processor

Dual Core

7 µm / 5 µm

27 µm / 19 µm

Processor Speed

2.5 GHz

Axial/Depth Resolution (Air/Tissue) Imaging Depth Range (Air) A-Scan/Line Rate a

1.7 mm

6.5 mm 1.25 kHz

Base Unit Dimensions 420 mm x 320 mm x 150 mm Using a Common Path Probe with 50% Beamsplitter

COMPUTER SPECIFICATIONSb

Memory Hard Drive Data Acquisition

Windows 7, 64 Bit

4 GB 250 GB USB

Computer Specifications Subject to Change

The Options OCTH-LK20-BB General Purpose Hand-Held Probe

Scan Lens Kit

OCTH-LK30-BB Long Depth of Focus

Reference Arm Adapter (Rigid Scanner Only)

OCT-LK2-BB High Resolution

OCT-RA2 Adapter

Rigid & Adjustable Cage-Compatible Scanners OCT-LK3-BB General Purpose

OCT-RA3 Adapter

Step 3:

Choose a Scan Lens Kit

OCT-LK4-BB Long Depth of Focus

Step 4:

CAL224 The CAL224 configuration consists of a long-range high-sensitivity 830 nm base unit, adjustable cage-compatible scanner, long-depth-of-focus scan lens kit, adjustable immersion Z-spacer, stand, and stage.

OCT-RA4 Adapter

Hand-Held Probe (Ring)

Choose a Z-Spacer (Optional)

OCTH-AIR20 for OCTH-LK20-BB Kit

OCTH-AIR30 for OCTH-LK30-BB Kit

Rigid & Adjustable Cage-Compatible Scanners

Step 5:

OCT-AIR3 for OCT-LK3-BB Kit (Ring)

Choose Accessories (Optional)

OCT-XYR1 XY Translation and Rotation Stage

OCT-PEDAL2 Foot Switch

OCT-STAND Stand for Rigid and Adjustable Cage-Compatible Scanners

OCT-IMM3 for OCT-LK3-BB Kit (Immersion)

OCT-IMM4 for OCT-LK4-BB Kit (Immersion)

Imaging Systems Bergamo Series Microscopes

Custom OCT Systems

Cerna Series Microscopes

2D and 3D Imaging Solutions

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Thorlabs offers engineering solutions for integrating customized Optical Coherence Tomography (OCT) imaging modules into complex host systems. Our OCT groups consist of scientists and engineers with over 10 years of experience in OCT technology and its applications. Both systems and components are manufactured at our facilities in Lübeck, Germany, and Newton, NJ. We look forward to working with you to design a custom system that meets your needs.

OEM OCT System for Ophthalmology

Project Considerations To successfully integrate advanced imaging technology into a complex host system, there are many factors to consider beyond component development: • Selection of the best-suited parameters for the imaging technology within the context of the host system and its application • A dedicated engineering team that is in close contact with the host system development team • Customization of certain components for seamless host system integration • A reliable manufacturing process that provides timely delivery of high-quality components

Competencies n Extensive

Knowledge in Photonics Technology n Vertically Integrated Manufacturing Facilities n Over a Decade of Experience Developing OCT Technologies n Library of Existing OCT Systems, Components, and Subassemblies n Engineering, Customization, and Manufacturing Expertise

Experienced Team n Contributions

to the OCT Field for Over 10 Years n Extensive Portfolio of OCT Components Developed and Manufactured by Thorlabs n Several Hundred Systems Successfully Designed, Manufactured, and Deployed into the Field n Over 20 Peer-Reviewed Publications Involving Successful Research Conducted with Thorlabs’ OCT Systems and Components

0.5 mm 20 ms Healthy Human Lens Imaged with a Custom 1070 nm OCT System

114

2 mm 40 ms Anterior Chamber Imaged with a Custom 1070 nm OCT System

Imaging Systems Bergamo Series Microscopes

Custom OCT Systems

Cerna Series Microscopes

Engineering Process Thorlabs is committed to working with you at each stage of the development process to ensure the final design meets your unique requirements. 1

Process Analysis

Imaging

System

Feasibility Evaluation

OCT as Imaging Technology

Proof of Principle

Rapid Testing with Existing Components

Demand Identification

Component Requirements, Parameters, and Interfaces

Module Engineering & Testing

Customized Prototype

Manufacturing Process & Quality Control

Imaging Component for System Integration

Application Market

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Thorlabsâ&#x20AC;&#x2122; Expertise Thorlabs is a leading photonics company that develops and manufactures a broad portfolio of technologies ranging from optical components to advanced imaging systems. We have implemented OCT imaging solutions for a wide range of applications in various fields with different levels of customization.

Customized Probe Module to Allow Submerged Imaging of Liquids Customized OCT Probe, or Sample Head, Designed to Allow Imaging of Liquid within a Bottle

0.5 mm Contact Lens Imaged in Saline Inside the Transport Blister

0.5 mm Laminated Foil Measurement Imaged at 2 Îźm Axial Resolution

115

Imaging Systems Bergamo Series Microscopes

TDI Whole-Slide-Scanning Microscope

Cerna Series Microscopes

Microscope

Essentials Kit Confocal Microscopes ThorImageLS Software

PC with TDI Software Fast XY Scanning Stage with Precision Encoder

OCT Systems TDI Digital Microscopy Optical Tweezers

Scientific-Grade CCD Camera

TDI Engine

Features

Joystick

High-Speed Scanning of Whole Slides • Image 10 mm x 10 mm at 15X* in 35 Seconds • Image 10 mm x 10 mm at 31X* in 94 Seconds n Exposures from 5 ms to 250 ms Suitable for Brightfield and Fluorescence Microscopy n Monochrome and Color Imaging Available n Compatible with Many Commercially Available Microscopes n Use Whole Slide Scans to Quickly Identify Areas of Interest for Further Analysis • GUI to Select Areas of Interest and Move the Stage to that Precise Location • Switch to Standard Video Imaging Mode for Further Inspection and Analysis n

*Magnification Calculated Based on DICOM Standard with 10X and 20X Objectives

116

Scientific Challenge

Solution

Conventional whole-slide imaging methods employ a “stop-and-stare” approach, which requires the system to accelerate, step across a sample, decelerate, stop, and settle at each location to acquire an image. As researchers seek to acquire whole-slide images as part of their everyday workflow, the throughput of conventional scanning systems presents a significant performance bottleneck.

Thorlabs’ Time Delay Integration (TDI) electronically eliminates the relative motion between the sample and the imaging array, allowing an image to be acquired without stopping the motion of the slide, even for long exposures.

Imaging Systems

TDI Whole-Slide-Scanning Microscope Thorlabs’ TDI Whole-Slide-Scanning System provides a fast, accurate solution for whole-slide imaging. This patent-pending technique synchronizes the position of the stage with the transfer of charges across the camera’s CCD sensor to effectively eliminate the relative motion between the sample and the imaging array. Doing so enables longer

effective exposures without stopping motion, while also eliminating image alignment errors inherent in stop-and-stare imaging. An additional benefit is the significant increase in scanning throughput. See the table on page 118 for a comparison of TDI and Stopand-Stare methods for similar exposures.

Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

A TDI image of a tumor in lung tissue taken at 31X* magnification. The inset view shows a small area of the whole-slide image to illustrate the level of clarity and detail. Scan area: 12 mm x 20 mm. *Magnification Calculated Based on DICOM Standard with 20X Objective

Perform Whole-Slide and Detailed Studies

Once the whole-slide image is acquired, the software allows the user to enlarge areas of particular interest. When a region of interest is identified by the user, the stage can be moved to that position and the

camera switched to standard video mode. This allows for detailed study with the desired microscope and camera imaging techniques. For example, the sample could be viewed at higher magnifications.

A TDI image of a mouse kidney stained with AlexaFluor® 488. The inset view shows a small area of the whole-side image to illustrate the level of clarity and detail.

117

Imaging Systems Bergamo Series Microscopes Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

The Advantages of TDI Whole-Slide Scanning Speed

SCAN TIME

The advantages of TDI increase STOP AND THROUGHPUT AREA MAGNIFICATION* STARE TDI IMPROVEMENT as magnification increases. By 10 mm x 10 mm 15X 1 min 32 sec 35 sec 260% taking advantage of the control 10 mm x 10 mm 31X 6 min 30 sec 1 min 34 sec 318% loop feedback, the Thorlabs TDI 25 mm x 75 mm 15X 18 min 5 min 50 sec 308% system produces images with high 25 mm x 75 mm 31X 86 min 23 min 373% positional accuracy at significantly *Magnification Calculated Based on DICOM Standard with 20X Objective faster speeds than stop-and-stare imaging methods. Some typical examples of slide scan times using a standard stop-and-stare system versus Thorlabsâ&#x20AC;&#x2122; patent-pending TDI technology are outlined in the table above.

Higher-Quality, Large-Format Images Thorlabsâ&#x20AC;&#x2122; TDI system is ideal for applications where positional accuracy is paramount. The integration of the camera into the motion control loop allows for precise image registration at the pixel level while capitalizing on the speed of the stage. This unique implementation can provide better positional information with respect to the image than standard stop-and-stare methods. High positional accuracy is achieved by embedding positional information in the image file, allowing the rows of pixels to be tied to absolute positions that are used to create a large tiled image. Unlike algorithms employed in traditional stitching methods, which align each frame using image features, TDI creates the image based on absolute position. Thus, the risk of lost data that is inherent in many standard stitching schemes is not a concern using TDI. In addition, the TDI scheme allows for large-format imaging with high position image alignment for samples with sparse features. This is typically a challenge with off-the-shelf stitching programs that rely on feature recognition to align images.

A color TDI image of a Tiki Goddess, a trichrome-stained frontal young mouse tissue section taken at 15X* magnification. The inset view shows a small area of the whole-slide image to illustrate the clarity and level of detail. Sample courtesy of Dr. George McNamara, MD Anderson Cancer Center, Houston, Texas. Note: the image has not been retouched. Scan Area: 25 mm x 70 mm *Magnification Calculated Based on DICOM Standard with 10X Objective

118

Imaging Systems Bergamo Series Microscopes

The Advantages of TDI Whole-Slide Scanning Image Output The included TDI software provides a user-friendly viewer, which allows seamless zooming of the high-resolution images and the ability to select and save certain regions of interest. The images also provide positional information that can be used to relocate the microscope stage. Select the relocate mode in the software, and then click on the area of the image where you wish the stage to move. This feature is useful for live viewing of a certain region of interest at higher magnification or when using third-party software since it allows remote control of the microscope stage and camera parameters using the large-format image as a map. In addition to composite large-format images, the TDI system can also save each high-resolution image in tiff or jpeg format. These file formats provide full flexibility for any post processing required. Furthermore, each image includes meta data to relocate the translation stage for post-scanning live view, an ideal option for those wishing to revisit and review a region of interest in live image mode. The file structure also allows the user to either save the entire data set or only save regions of interest. This reduces the amount of data saved and improves the ease with which results can be shared with remote collaborators.

Cerna Series Microscopes

Have you seen our...

Scientific-Grade CCD Cameras ◆ 1.4, 4, or 8 Megapixel as well as ◆ ◆ ◆ ◆

Fast Frame Rate Cameras Available Low Read Noise at 20 MHz Readout Asynchronous, Triggered, and Gated Exposure Modes ThorCam GUI Third-Party Software Support Including LabVIEW, MATLAB, μManager/ImageJ, and Metamorph

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

1500M-GE-TE

See Pages 308 - 322

A screenshot of Thorlabs’ TDI software. The whole slide scan of the lung tissue section is shown on the left side of the main window. The objective icon indicates the area seen in the main view to the right. By moving the objective icon with the mouse or by moving the joystick, the area shown in the main view will update accordingly. From this screen, specific regions of interest can be selected for further study with other imaging techniques. 119

Imaging Systems Bergamo Series Microscopes

TDI System Components

Cerna Series Microscopes

Thorlabsâ&#x20AC;&#x2122; TDI system is available as a fully configured item, including the camera, microscope, stage, TDI engine, and PC. Alternatively, the system may be adapted to fit a microscope already installed in your lab. Please contact Sales.tsi@thorlabs.com to discuss your system configuration.

Essentials Kit Confocal Microscopes

Microscope n Brightfield

and/or Fluorescence Imaging n Compatible Objectives: 4X, 10X, 20X, and 40X n Microscope Included (Call for Available Configurations) n Available without Microscope for Integration into Existing Lab Setups (See Table Below for a List of Supported Microscopes)

ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

MLS209-1

TDI System on an Nikon Microscope

Stage Shown with an MLS203P9 Slide Holder

High-Speed Precision Stage with Encoder n Encoder

Generates Precise Position Information n Linear Motors Provide Smooth Motion for Improved Accuracy n Single or Multiple Slide Holders Specifically Designed for TDI

Scientific-Grade Interline CCD Camera n Low

Noise, High Sensitivity n GigE or Camera Link Interface

SUPPORTED MICROSCOPES

120

Upright

Inverted

Olympus BX41, BX43, BX51, BX53, and BX61 Nikon Eclipse FN1 Nikon 50i, 80i, 90i, and Ci-L

Olympus IX81, IX73, and IX81 Nikon Eclipse Ti Nikon TE2000 Zeiss Axio Observer and Zeiss Axiovert 40

Imaging Systems Bergamo Series Microscopes

TDI System Components TDI Software

TDI Engine

n Full-Featured

Windows 7 GUI for TDI Acquisition n Intuitive Setup of Whole-Slide-Scanning Parameters n Save-to-Disk and Visualization of Whole-Slide Images n Interactive Zoom In/Out Capabilities with Inset View n Right Click to Move Stage to Region of Interest within Whole-Slide Image

n Controls

Movement of the Stage During TDI as well as Conventional Imaging n Performs All Computation and Synchronization Necessary for TDI Operation n Connection Cables for Stage, Camera, and PC Included n Control via TDI Software on PC

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

PC with TDI Software Included

MJC001 Joystick

TDI Engine

Joystick n Enables

Efficient XY Location of Home Position as well as Areas of Interest n Speed Adjustment for Fast or High Precision Moves n Speed Dial for Sensitivity Adjustment n Ergonomic Design Ensures Quick, Tactile, and Interactive Workflow n High-Quality Machined and Anodized Aluminum Casing n High-Reliability Hall Effect Joystick

Computer,* Monitor, Keyboard, and Mouse n Fully

Configured with TDI Software and Drivers to Communicate with the TDI Engine and Scientific Camera n Optimized for Whole-Slide-Scanning Workflow *PC Model and Configuration Subject to Change

ITEM # T1500M-GE-KIT

PRICE 36,000.00

DESCRIPTION Brightfield TDI Upgrade System for Existing Microscopes

T1500M-GE-NIKU

56,000.00

Brightfield TDI System with Nikon Microscope

The TDI System is offered in several varieties with prices starting at $36,000. Please email us at Sales.tsi@thorlabs.com or call us at 973-300-3000 to discuss the various options. 121

Imaging Systems Bergamo Series Microscopes

How it Works: Time Delay Integration Microscopy

Cerna Series Microscopes

In TDI, the charge pattern on the CCD is accumulated and shifted while moving the sample through the field of view of the microscope. The user starts by entering a “home,” or starting position, and the region of interest in the TDI software. The stage moves past the objective, changing the field of view seen by the camera. The encoder in the stage sends signals to the TDI engine, allowing it to track the position of the stage.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

As the sample moves through the field of view of the microscope, a charge pattern is accumulated on the CCD.

One “Strip” of a TDI Scan

8 8 8 3 3 3 4 4 4

Third Strip of TDI Scan Starts Here Second Strip of TDI Scan Starts Here “Home” First Strip of TDI Scan Starts Here

Direction of Stage Motion

The region of interest is divided into strips as shown here. The “home” position is in the lower right hand corner.

The region of interest is divided into strips to determine the scanning pattern. The schematic above shows an example of a slide divided into strips, each one indicated by a different color. Although we will only look at three strips for this example, the actual number of TDI strips is calculated to accommodate the entire slide at the desired magnification. The program will begin the scan at the home position. Images from each strip are used by the TDI software to create the full image of the slide. The picture to the right shows the TDI system in the "Home" position, before the user has pressed the "Start" button in the TDI software.

TDI System Configuration Prior to Acquiring Data

122

Imaging Systems Bergamo Series Microscopes

How it Works: Time Delay Integration Microscopy

Cerna Series Microscopes

When the field of view moves by the equivalent of a pixel row, the TDI engine will trigger the camera to shift the accumulated charge to the next row. When the row of charge reaches the edge of the CCD chip, it is read out to the computer. Charges are accumulated in the direction of the scan, increasing the effective exposure.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers TDI System Scanning Across the First Strip of the Slide

Once the first strip of the slide is scanned, the stage will move to its starting x-position and shift the y-position by the width of the first strip. It will then begin to scan the next strip of the slide.

TDI System Scanning Across the Second Strip of the Slide

In this way, the entire slide can be quickly scanned while moving the stage and without introducing blur. After the scan is finished, a complete image, comprised of all of the strips, is displayed in the imaging software. This image can be zoomed in on and examined to identify features that require further examination. A simple go-to command in the software package allows the user to send the stage to these selected areas for further analysis using higher magnification or other imaging conditions.

The Last Strip of the Slide is Scanned and Read Out to Complete the Whole-Slide Image

For an animation that describes the operation of TDI, please visit www.thorlabs.com/WholeSlideScanning. Please contact Sales.tsi@thorlabs.com for pricing and availability. 123

Imaging Systems Bergamo Series Microscopes

Optical Tweezers Overview

Cerna Series Microscopes

Optical Tweezers have become an important tool in a wide range of fields such as bioengineering, materials science, and physics due to their ability to hold and manipulate micron-sized particles and to measure forces in the piconewton range. Applications include trapping viruses and bacteria, manipulating cellular structures, patterning of surfaces, integration with fluorescence imaging, and measuring force of molecular motors and biological molecules such as DNA and proteins.

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Thorlabs offers two separate optical tweezers systems (shown below), each with its own distinct advantages. The Microscope Optical Tweezers provide a fully assembled optical tweezers solution, while our Modular Optical Tweezers are ideal for those seeking a setup with an open architecture.

OTM200 Microscope Optical Tweezers Our OTM200 Microscope Optical Tweezers System (shown to the right) was created for scientists who desire an out-of-the-box, optical tweezers solution for inverted microscopes. This system is available with a Nikon Eclipse Ti microscope, a high-resolution XYZ piezo-driven stage, a laser source, control and data acquisition electronics, and a computer with pre-installed software. The OTM200 can also be adapted to connect to a variety of ports on inverted microscopes from all major manufacturers. OTM200 Microscope Optical Tweezers System (For Complete System Details, See Pages 126 - 131)

OTKB Modular Optical Tweezers

OTKB Modular Optical Tweezers (For Complete System Details, See Pages 132 - 135)

The OTKB Modular Optical Tweezers (see pages 132 - 133 for details) consists of five pre-assembled segments used to construct the optical tweezers system. The modularity of this arrangement allows users to customize, modify, or upgrade the system with standard components as needs change and provides more flexibility over other closed optical tweezers systems. Three application modules are also offered that are designed to enhance the capabilities of the Modular Optical Tweezers (see pages 134 - 136).

Accessories Also available is a sample preparation kit to quickly prepare a sample with Ă&#x2DC;1 Âľm fused silica beads and to test any optical tweezers system. High-numerical-aperture objectives are also available for applications that involve the optical trapping and manipulation of higher index particles. 124

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Optical Tweezers Theory

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Optical Tweezers are created by using a high-numericalaperture objective to tightly focus a laser beam, creating an area where a micrometer-sized particle will experience a force due to the transfer of momentum from scattering photons. Two regimes are considered when calculating forces exerted by an optical trap on a particle. If the trapping wavelength is less than the diameter of the particle, the Mie scattering theory is employed. If the wavelength is greater than the particle diameter, the Rayleigh scattering treatment is considered.

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Mie Regime

Optical Tweezers

The Mie regimea can be described using ray optics. When laser light, which normally has a Gaussian intensity profile, interacts with a dielectric particle, it gets refracted and reflected. It is important to note that the momentum of the light will change when either of these mechanisms changes the direction of light propagation. Conservation of momentum requires that the particle must have an equal and opposite momentum change, resulting in forces acting on it. Refraction creates a force that draws a particle towards the center of the beam. Reflection, on the other hand, creates a scattering force along the direction of light propagation. In the case where a particle is not aligned axially in the center of the laser beam, as shown in Figure 1 with a collimated beam, the refracted rays closer to the center of the beam will be more intense and will transfer more momentum to the particle. This applies a lateral restoring force to the particle towards the center of the beam. Once the particle is in the center of the beam, the rays refracting through the particle will be symmetric, as shown in Figure 2 with a focused beam, and the particle will be laterally trapped. Unless there is a steep gradient of light intensity, the scattering force will push the object out of the trap. This gradient is obtained by using a highnumerical-aperture objective, which produces a gradient force large enough to overcome the scattering force, and as such, the trap location will always be offset from the focal point of the objective.

Rayleigh Regime In the Rayleigh regime, the diameter of the particle is much smaller than the wavelength and the ray theory breaks down. To understand the forces, the trapped particle is considered a point particle. The scattering force arises from absorption and re-radiation, while the gradient force results from the interaction between the inhomogeneous electric field of the laser beam and the induced dipole moment of the particle.b In most situations, however, the particle sizes are comparable to the wavelength. The complete treatment becomes too complex to discuss here but has been described in several publications.c

Laser In Laser In 1

Objective Lens 2

Fgradient F1

Fscattering

Fscattering Figure 1. Particle being restored to the center of the trap in a collimated laser beam

Figure 2. Particle held in equilibrium within a focused laser beam

A. Askin et al. Opt. Lett. 11, No. 5, 288, 1986 C. N. Keir and M. B. Steven. Review of Scientific Instruments 75, No. 9, 2004 c E. Almaas and I. Brevik. J. Opt. Soc. Am. B 12, 2429, 1995; P. Zemanek et al. J. Opt. Soc. Am. A 19, 1025, 2002; K. F. Ren et al. Opt. Commun. 108, 343, 1994 b

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Microscope Optical Tweezers System OTM200

Features

Microscope Tweezers System Integrated with an Olympus IX83 Inverted Microscope (Force Module Not Shown)

n Complete

Optical Trapping System n Available with a Nikon Eclipse Ti Microscope (Optical Integration with Other Major Inverted Microscope Brands Available) n Multiple Computer-Controlled Traps n 1064 nm Laser Source (Other Trapping Wavelengths Available Upon Request) n Force Measurement and Particle Tracking

The OTM200 is a complete out-of-the-box optical tweezers system for an inverted microscope. It can operate in conjunction with other imaging modalities such as confocal microscopy and Raman spectroscopy. The trapping source is a power- and wavelength-stabilized 1064 nm laser, which is split into two independently steerable trapping beams, each capable of supporting several time-shared traps. Other trapping wavelengths are available for specialized applications. A GUI control software provides plug-and-play support for most general trapping experiments. In addition, a software development kit enables users to create application-specific solutions. The optical tweezers software package also includes support for high-end EMCCD cameras to enable the combination of optical trapping and fluorescence imaging in light-starved applications. An Andor low-noise EMCCD camera is available as a custom order. The OTM200 Optical Tweezers System is available with a Nikon Eclipse Ti microscope, a high-resolution XYZ piezodriven stage, the laser source, control and data acquisition electronics, and a computer with pre-installed software as shown below. It can also be adapted to connect to a variety of ports on inverted microscopes from all major manufacturers, as shown above. Please contact applications@thorlabs.com for more information about customization or integration with an existing microscope. The system conveniently fits on a 3' x 4' (900 mm x 1200 mm) ScienceDeskTM Workstation with active vibration isolation (not included), as shown in the photo to the right. See pages 364 - 365 to learn more about our ScienceDeskTM frames, tabletops, and accessories for use with our optical tweezers system. A sample table layout is given on page 131.

The OTM200 Tweezers System shown with a Nikon Eclipse Ti microscope, a high-resolution XYZ piezo-driven stage, the laser source, control and data acquisition electronics, and a computer with pre-installed software

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Applications n Trapping

Viruses and Bacteria n Manipulation of Cellular Structures n Patterning of Surfaces n Integration with Fluorescence Imaging n Force Measurement of Molecular Motors and Biological Molecules such as DNA and Proteins

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Microscope Optical Tweezers System

Cerna Series Microscopes

Software The OTM200 system includes a Windows-based software package that contains everything needed for system control and data acquisition. The GUI control software provides plug and play support for most general trapping experiments (shown in Figure 1). In addition, a software development kit enables users to create application-specific solutions. The tweezers system also includes support for high-end EMCCD cameras to enable the combination of optical trapping and fluorescence imaging in light-starved application. An Andor lownoise EMCCD camera is available as a custom order. For more information about the software used in the Microscope Optical Tweezers, please visit our website.

Software n Trap

Positioning via On-Screen Manipulation (Video Available at www.thorlabs.com) n Adding / Removing Trap Sites n Stage and Trap Step Positioning n Laser Power Control n Camera Control

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Software Development Kit The tweezers system is supplied with a Software Development Kit (SDK). The SDK gives access to all features of the instrument, thus enabling the creation of custom, application-specific software. The SDK is provided as a 64-bit Windows dynamic link library (DLL). Language bindings for C, LabVIEW, and C# are available. Figure 1. The main screen of the OTM200 software is shown with three Ø1 µm beads trapped. They are labeled A_1, A_2, and B_1.

Force Measurement The optional force measurement module allows users to apply and measure forces in the piconewton range. The tweezers system allows the user to run an automated calibration sequence in situ without the need for a special sample, which might not match the condition during the experiment. Additionally, the position of the particle can be tracked in three dimensions. Figure 2 shows a histogram of a trapped particle’s position in the proximity of a glass surface. The XY data is symmetric, whereas the Z-data is truncated, as the particle cannot penetrate the surface. Figure 3 shows the software screen showing the position and force tracking as measured by the force module.

Figure 2. A histogram showing a trapped particle’s position in three dimensions in proximity to a glass surface.

Figure 3. Software screen showing position and force tracking.

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Microscope Optical Tweezers System

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Simultaneous Trapping of Multiple Cells

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Optical tweezers can be employed to trap biological cells. While the cells themselves can be quite large and are filled mostly with water, the tweezers can trap the cell nucleus or other organelles, thereby stabilizing the cell’s position. It is also possible to attach functionalized polystyrene particles to the cell walls and use them as “handles.” Figure 4 shows live yeast cells in a microfluidic channel. In the absence of flow, the cells have sedimented on the bottom of the channel. The OTM200 tweezers system has picked up seven cells from the channel bottom and simultaneously holds them in a circle in the focal plane.

Figure 4. OTM200 Optical Tweezers System holding multiple yeast cells simultaneously.

Combination with Other Imaging Modalities Thorlabs’ Optical Tweezers System can be combined with a wide variety of imaging and spectroscopic methods, such as phase contrast, differential interference contrast, fluorescence imaging, confocal imaging, or Raman spectroscopy. Figure 5 shows a combination of optical trapping and fluorescence microscopy. A Ø15 µm polystyrene sphere was dyed using Dragon Green and is being lifted by the optical tweezers in the location Figure 5. Combination of optical trapping and labeled “Trap A 1.” In the adjacent trap, labeled “Trap A 2,” a fluorescence microscopy. Ø2 µm sphere that has not been dyed is being held. The dye was excited using a high-power plasma light source, which was coupled into the microscopy path using a standard filter set (see pages 228 - 231 for more details on the filters).

DNA Stretching

Force (pN)

Thorlabs’ OTM200 Optical Tweezers can be used to measure the forces within DNA. As shown in the inset in Figure 6, one end of a 1 μm long piece of dsDNA was attached to a glass slide and the other end was attached to a Ø1 μm polystyrene bead. While the OTM200 was used to hold the bead, the microscope stage was translated along its Force Acting on Bead x-axis at a rate of 10 μm/s, stretching the DNA. DNA

Laser Beam

The displacement (dx) of the bead was measured using the OTM200 force module, and the force was calculated using F = -k•dx, where k is the stiffness of the optical trap. Figure 6 illustrates a typical force versus extension curve obtained for bare DNA, represented by the red circles, with error bars denoting the standard deviation.

1.0

When stretched with a force of 65 pN, the DNA demonstrates a phenomenon known as an overstretching transition, which can be observed in Figure 6. The sharp decrease in force seen at the end of the plot was caused by the bead escaping from the optical trap and being pulled back to its original position by the attached DNA.

Overstreching Transition

Extension 40 Glass Surface 20

0 0.0

0.5

1.5

2.0

Extension (µm) Figure 6. A typical force vs. extension curve for bare DNA measured with the OTM200 Optical Tweezers. The error bars represent the standard deviation. The inset is a schematic showing a setup used to measure the forces within DNA.

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Microscope Optical Tweezers System

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Trapping of Metallic Nanoparticles and Single Molecules

Essentials Kit

Nanoparticles and single molecules can be trapped by optical tweezers under certain circumstances. Trapping of gold particles with diameters ranging from tens of nanometers to a few hundred nanometers has been reported. These nanoparticles play an important role in in vivo studies due to good biological compatibility. Please contact applications@thorlabs.com to discuss your samples.

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Figure 7. A diagram showing a setup for trapping Bovine serum The illustration in Figure 7 shows a specialized sample albumin (BSA) proteins. environment that has been implemented using Thorlabs’ tweezers. It utilizes plasmonic enhancement to trap gold nanoparticles with a diameter of 10 nm, as well as single bovine serum albumin (BSA) proteins. The BSA molecules have a hydrodynamic radius of 3.4 nm. For details, see Pang and Gordon (Nano Lett. 12, 402-406, 2012 and Nano Lett. 11, 3763–3767, 2011).

TDI Digital Microscopy Optical Tweezers

Microrheology Microrheology is the study of the flow of materials, including measurements of viscosity and viscoelasticity, over micron length scales. One common measurement is to look at the response of a microscopic object subject to an external force; optical tweezers can be employed to apply the external force while the movement of the particle is recorded. Figure 8 shows two measurement schemes using optical tweezers: either an organelle inside the cell can be trapped for use as a probe or the probe can be a silica particle attached to the cell’s membrane.

Silica Particle

Cell Organelle

Nucleus

Cell Membrane

Figure 8. Two Microrheology methods using optical tweezers. Left: organelle inside the cell is trapped and is used as a probe for measurements. Right: probe is a silica particle attached to the cell’s membrane.

Passive Microrheology Near a Cell Wall 1E-6

Bead Above Cover Glass

Position Power Spectral Density (V2/Hz)

Bead 12 µm Above Cell Wall Bead 7 µm Above Cell Wall

1E-7

Bead 2 µm Above Cell Wall Bead Stuck on Cell Wall

1E-8

1E-9

1E-10

1E-11 20

100

1000

Frequency (Hz) Figure 9. Plot showing the position PSD as a fused silica bead is moved closer to the cell wall

6000

Figure 9 shows the position power spectral density (PSD) of a particle located just above a coverslip as well as at 5 different distances from a cell wall. The PSD shows the frequency content of the particle’s motion. It can be seen that the higher frequency components of the particle’s motion are suppressed. A particle in a Newtonian fluid will have a PSD with a Lorentzian shape (refer to the blue and red curves in the figure). As the particle is moved closer to the cell wall, the PSD begins to deviate from the Newtonian ideal and more complex models must be used to describe the fluid’s behavior. 129

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Microscope Optical Tweezers System

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Integration with Microfluidics

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Many applications in biochemistry and biophysics require the addition and exchange of reagents over the course of an experiment. Microfluidic and “lab-on-a-chip” devices allow for the exchange of buffers, the addition of reagents, and the creation of reagent gradients with high speed and precision. Fluid flow through a microfluidic device can also be utilized to apply well defined viscous drag forces (e.g., to extend single DNA molecules). The OTM200 Microscope Tweezers have been successfully integrated with commercially available and customized flow cells. A simple, 100 μL flow channel is sold as part of the OTKBTK Sample Preparation Kit (see page 137) and is shown in Figure 10.

Figure 10. OTM200 Microscope Tweezers shown using a 100 µL flow channel from the OTKBTK sample preparation kit. Also shown are supply and exaust lines, which are connected to a syringe pump (Not Shown).

For cell sorting, a suitably designed microfluidic device can be used in conjunction with the OTM200 Microscope Tweezers to sort particles. Cells can be held in a reservoir, where they are trapped and evaluated using fluorescence or Raman spectroscopy, for example. Based on the result, the tweezers can then transport the cells of interest into a certain flow channel or reservoir for safekeeping and further processing. The remaining cells can then be discarded.

Integration with EMCCD Cameras The OTM200 Optical Tweezers System software package includes support for EMCCD cameras, which are ideal for low-light applications. An Andor EMCCD camera is available as an option for the OTM200 optical tweezer system. An example of a study enabled by the EMCCD is shown in Figures 11 and 12 below, which display results from a study of a kinesin motor moving on a microtubule, a component of the cell’s cytoskeleton. A Ø1 µm polystyrene bead with a kinesin motor attached is brought close to a microtubule; the trap is then held stationary and the bead and motor are latched onto the microtubule by Brownian motion. Once attached, the kinesin motor attempts to move along the microtubule, which displaces the bead from the center of the trap. This displacement is measured with the tweezers’ force module, and the results are plotted in Figure 11. In the region indicated in green in the plot, the bead and motor move two distinct, 8 nm steps, indicating the size of the tubulin proteins are 8 nm. Figure 12 shows the rhodamine-labeled microtubules (excitation peak: 535 nm, emission peak: 585 nm) and the polystyrene bead viewed with an EMCCD camera, as well as a schematic of the experiment. The light levels are too low to locate the position of the microtubules with a standard camera. 0.02 Tubulibn Dimer (8 nm)

0.00

dx (µm)

8 nm 8 nm

Microtubule

-0.02 Bead Kinesin Motor

-0.04

1000

2000

3000

Time (ms) Figure 11. The displacement of a trapped Ø1 µm bead with a kinesin motor attached as the motor moves along a microtubule.

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Figure 12. Right: An image taken with an EMCCD camera of a Ø1 µm bead with a microtubule. Left: A schematic of the kinesin motor, with attached microbead, moving along a microtubule.

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Microscope Optical Tweezers System Trap Functionality The output of the 1064 nm trapping source for the Microscope Optical Tweezers is collimated, and the light is focused onto the sample with diffraction-limited performance, thereby achieving trapping forces exceeding 200 pN. The trapping laser is OPTICAL TRAP SPECIFICATIONS capable of providing two independent Center Wavelength 1064 nm trapping beams, which can be precisely Emission Bandwidth <0.25 nm positioned in three dimensions. The stiffness Power per Trap Beama >1 W of each trap can be individually controlled Beam Diameter at Back Aperture 8 mm and is actively stabilized. Number of Independent Trap Beam Paths 2 Maximum Number of Trap Sites per Beam Pathb

Areab

> 80 µm x 80 µm

Laser Power Noise (RMS)a

<0.2%

Trap Scan

MICROSCOPE SPECIFICATIONS Example Inverted Microscopes Focusing

Nikon Eclipse Ti Olympus IX Series Via Nosepiece, Manual or Motorized

Condenser

0.9 NA

Objective

>1.2 NA, Oil Immersion

200 µm XYZ Travel Piezo Stage, 1 nm Resolution

Detection

Dual QPD Module

Data Acquisition

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System Features

OPTIONAL FORCE MODULE Sample Stage

The table to the left lists the specifications of the optical tweezers as well as common options for configuring a complete solution. For more information about our optical tweezers, or for assistance in building a system that meets your needs, please contact us at applications@thorlabs.com.

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16 Bits, 5 ns Timing Resolution

Measured at the back aperture of the objective on a Nikon Eclipse Ti microscope. b Tested with Ø2 µm fused silica beads in water, using an RMS100X-PFO objective (100X Olympus Plan Fluorite Oil Immersion Objective, 0.2 mm WD), and a No. 1.5 coverslip on a Nikon Eclipse Ti microscope. c Please contact applications@thorlabs.com with questions regarding integration with other inverted microscopes or for information about purchasing a system with a microscope.

Multiple Computer-Controlled Traps n Various Wavelength Options Available • 830 nm • 976 nm • 1064 nm n Integrates with Existing Inverted Microscope n Software Package Including SDK n

Suggested ScienceDesk™ Configuration 1 Nikon Eclipse Ti-S Microscope 2 Optics Module (Included with OTM200) 3 Power Supply and Control Modules (Stacked, Included with OTM200) 4 Nexus™ Tabletop (Not Included with OTM200, See Page 364) 5 SDA90120 3' x 4' ScienceDesk Workstation (Not Included with OTM200, See Page 364) 6 PSY351: Side Shelf (Not Included with OTM200, See Page 365)

ITEM # OTM200*

PRICE $ 82,000.00

DESCRIPTION Optical Tweezers Upgrade Configuration for Existing Microscopes

*Please contact applications@thorlabs.com for more information regarding the purchase of a system with the optional force module and/or a system including an inverted microscope.

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Modular Optical Tweezers

Cerna Series Microscopes

n Complete

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems TDI Digital Microscopy Optical Tweezers

Optical Trapping System n Inverted Light Microscope Design Ideal for In Vitro Biological Experiments n 975 nm, 340 mW Trap Laser with Easy-to-Use Touch Screen Controller n Nikon 100X Oil Immersion Objective Thorlabsâ&#x20AC;&#x2122; OTKB Modular Optical Tweezers have the flexibility required for experiments conducted in advanced research laboratories. The modular system is based on an inverted microscope design that provides compatibility with standard samples. It is offered with a CMOS camera for video imaging, and the system can easily be adapted for fluorescence, single-molecule, and other types of microscopy. The advantage to purchasing and assembling this modular

n 3-Axis

Sample Positioning Stage with Integrated Piezo Actuators for Nanometer Resolution n CMOS Camera with USB Interface for Video Imaging n Force Measurement and Fluorescence Microscopy Modules Available system is the versatility it provides over other closed optical tweezers systems. Since the optical tweezers are built using standard Thorlabs components, it is easy to modify or upgrade the system using other standard components. For example, the system can be easily modified to produce multiple trapping beams or to enable the oscillation and/or steering of the tweezersâ&#x20AC;&#x2122; position using a galvo mirror.

White-Light LED OTKBFM Back Focal Plane Detection Module (Optional)

Have you seen our... Application Modules

Controllers for Stage and OTKBFM Quadrant Position Detector

OTKB-FL Fluorescence Module (Optional) Condenser Sample Positioning Stage

Objective

Laser Diode and Controller

An Optional Force Measurement Module as well as other modules are available

See Pages 134-136 OTKB

Shown with Optional Fluorescence and Back Focal Plane Detection Modules

Camera Beam Expander

Fiber Collimator

The OTKB is shipped in preassembled segments that only require final coupling and alignment; however, we can ship a system assembled, aligned, and tested on a breadboard for an additional cost. Custom versions optimized for advanced teaching laboratories are available; for example, configurations with a lower power laser or a manual sample stage can be provided to reduce costs. For details, please contact applications@thorlabs.com. 132

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Modular Optical Tweezers

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System Description The 975 nm trapping laser source is a pigtailed Fiber Bragg Grating (FBG) stabilized single mode laser diode in a hermetically sealed 14-pin butterfly package. The integrated TEC element and thermistor in the butterfly package allow the temperature of the laser to be precisely controlled with a touch-screen laser diode / TEC controller and mount. This leads to very stable output power, which is important for maintaining a constant trapping force. The laser input is collimated using one of our triplet fiber collimators to provide excellent beam quality and ease of alignment. A 3X beam expander is used to fill the aperture of the focusing objective. The dichroic mirror reflects light (trapping source) into the vertical path of the setup where a 100X, oil-immersion objective lens with an NA of 1.25 is used to focus the trapping laser beam down to a spot size of 1.1 µm.

LD & TEC Controller and Butterfly Mount

White Light Source

Essentials Kit Confocal Microscopes

Position Sensing Detector Dichroic Mirror Lens

ThorImageLS Software

Condenser Lens

Fiber

Sample Holder

Fiber Collimator

TDI Digital Microscopy Optical Tweezers

Objective Lens XYZ Sample Stage Mirror

OCT Systems

Lens

CMOS Camera

Beam Expander Dichroic Mirror

Schematic of Modular Optical Tweezers The microscope slide is positioned using a 3-axis (X, Y, and Z) translation stage that provides 4 mm of manual travel in combination with 20 µm of piezo actuation and a resolution of 20 nm. Using the internal strain gauges for positional feedback, 5 nm resolution can be achieved. The complete sample holder setup is placed on a translating breadboard, which facilitates loading/unloading of samples. The trapping laser is collimated by the condenser and reflected down the optional OTKBFM Back Focal Plane Detection Module (see page 134). Visible light from the white-light LED source illuminates the sample and is then imaged on the 1280 x 1024 pixel color CMOS camera using an achromatic doublet. The dichroic mirror in the lightpath in combination with a shortpass filter prevents saturation of the CMOS sensor as a result of backscattered light from the trapping laser. With the camera, you can acquire still images or video in color or monochrome black and white.

System Modularity and Assembly The modular tweezers system is shipped in pre-assembled segments. These segments will only require final coupling and alignment on a user-provided optical table or breadboard (see pages 378 - 381 for optical tables). A step-by-step instruction manual is provided. Once constructed, we recommend our OTKBTK Sample Preparation Kit (see page 137), which provides users with everything necessary to prepare a sample and determine that their optical tweezers are aligned and are functioning. ITEM # OTKB

METRIC ITEM # OTKB/M

PRICE 16,900.00

OPTICAL TWEEZERS SPECIFICATIONS Trap Force

~10 pNa

Spot Size

1.1 µm

Depth of Focus

1 µm

Power at Fiber Output

340 mW (Max)

OBJECTIVE SPECIFICATIONS Type Numerical Aperture

Nikon 100X Oil Immersion Objective 1.25

Input Aperture

Ø5 mm

Working Distance

0.23 mm

Wavelength Range

380 - 1100 nm

Recommended Cover Glass Thickness

0.17 mm

CONDENSER LENS SPECIFICATIONS Type

Nikon 10X Air Condenser

Numerical Aperture

0.25

Working Distance

7 mm

Wavelength Range

380 - 1100 nm

a At

a trap wavelength of 975 nm, a beam diameter of 5.1 mm at the back aperture of the objective, and with 140 mW of laser power at the trap focus.

DESCRIPTION Modular Optical Tweezers – Base Module

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Modular Optical Tweezers: Application Modules

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Back Focal Plane Detection Module

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n Position

and Stiffness Calibration n Force Measurement n Simple Integration into OTKB Modular Optical Tweezers n Quadrant Position Detector and Strain Gauge Controller Software Included Thorlabs’ OTKBFM Back Focal Plane Detection Module offers the ability to calibrate the OTKB Modular Optical Tweezers system for position detection and measurement of small forces.

Force Measurement Calibration Two methods are used by the OTBKFM-CAL to calibrate the force measurements: Power Spectral Density (PSD) rolloff and Equipartition. In the PSD roll-off, the PSD of a time series of trapped particle positions (due to Brownian motion) is computed. This data is fit to the response of a harmonic oscillator with known damping due to the viscosity of the solvent. The PSD rolloff method offers a particularly effective way to discover an inaccurate position sensor detection calibration, since it does not depend on the Screenshot showing the PSD curve acquired during force calibration. detector responsivity. The equipartition method equates the average potential energy of the particle in the trap to the thermal energy of the solvent molecules. The potential energy is calculated from the time domain data recorded during the force calibration. Since both PSD roll-off and equipartition methods rely on different physical principles, the combined results provide a convenient way to verify the Screenshot showing the time domain data used calibration. in the equipartition force calibration method.

134

OTKBFM

For high-bandwidth / high-resolution measurements, a Quadrant Position Detector (QPD) can be placed in a plane conjugate to the back focal plane of the condenser. In that case, the signal generated by the QPD is sensitive to the relative displacement of the trapped particle from the trap center and can be used to determine the position, stiffness, and force of the optical tweezers. The optional OTKBFM module includes a QPD, optics, optomechanics, and controllers to add back focal plane detection to our OTKB modular tweezers setup. This module does not include any data acquisition (DAQ) hardware or software to capture and analyze the data, making it ideal for users who plan to use existing DAQ hardware and analysis software.

OTKBFM-CAL

Force Acquisition Module Users who would like an out-of-the-box solution to measure forces can use the OTKBFM Back Focal Plane Detection Module in combination with our OTKBFM-CAL Force Acquisition Module. The OTKBFM-CAL acquires the X, Y, and SUM channels from the QPD and uses a differential amplifier to allow fast stage positioning. The included software provides procedures for position calibration as well as PSD roll-off and equipartition calibration and force measurement acquisition.

ITEM # OTKBFM

PRICE $ 2,800.00

DESCRIPTION Back Focal Plane Detection Module for OTKB and OTKB/M

OTKBFM-CAL

$ 3,500.00

Force Acquisition Module for OTKBFM

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Modular Optical Tweezers: Application Modules

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Fluorescence Microscopy Module By combining one of our Fluorescence Microscopy Modules with our modular optical tweezers, researchers can visualize, manipulate, and rapidly characterize the properties of various samples and cellular structures. Such techniques can be used to detect the arrival of a single molecule into a small volume, detect the conformational changes of cellular structures or bacteria, study elastic properties of a single DNA molecule, demarcate different parts of a larger molecular complex, and measure the response of each to an applied force.

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OTKB-FL

TDI Digital Microscopy

Fluorescence imaging filters can be easily inserted or removed with the included DFM(/M) Quick-Change Filter Cube (see page 242). To allow users flexibility in choosing the best fluorophores for their application, an imaging filter set is not included with these fluorescence modules; Thorlabs does, however, offer fluorescence imaging filters for a variety of common fluorophores. See pages 228 -231 for details.

Optical Tweezers

Also recommended is a plan fluorite or semi-apochromatic objective such as the RMS100X-PFO or N100X-PFO (see page 224). The plan fluorite or semiapochromatic design provides a wider corrected wavelength range, which is ideal for fluorescence microscopy. These objectives have a higher numerical aperture than the standard objective included with the OTKB, allowing them to trap particles that are functionalized and typically used as a probe, such as polystyrene. The OTKB-FL(/M) Fluorescence Module is shipped from stock without a light source for users who wish to attach a user-supplied light source. A receptacle for a liquid light guide with a diameter of 3 mm is provided. Alternatively, the OTKB-FLS(/M) module is available as a make-to-order item complete with an X-Cite 200DC mercury vapor light source and a Ø3 mm liquid light guide. ITEM # OTKB-FL

METRIC ITEM # OTKB-FL/M

PRICE 1,616.00

OTKB-FLS

OTKB-FLS/M

8,100.00

Fluorescence Image of an actin network stained with rhodamine phalloidin (Excitation: 540 nm, Emission: 565 nm) acquired by our Modular Optical Tweezers with the optional Fluorescence Module.

DESCRIPTION Fluorescence Module for the OTKB Fluorescence Module Including Light Source for the OTKB

Application Example Consider a sample consisting of a diluted solution of Ø1.0 µm uniformly FITC-dyed polystyrene beads with an excitation wavelength of 480 nm and an emission wavelength of 520 nm. The excitation light is selected from the X-Cite 200DC mercury vapor light source using an MF475-35 excitation filter, which provides transmission in excess of 85% in the 470 - 490 nm range. The light is then coupled into the tweezers system using an MD499 dichroic mirror, which reflects light in the 470 - 490 nm range and transmits light in the 508 - 675 nm range, as shown in the schematic to the right. The fluorescence light emitted by the sample will be collected by the objective along with any reflected excitation light, which gives a better signal-to-noise ratio than a transmissive detection scheme. The signal then goes back through the dichroics and an MF530-43 emission filter, which has a center wavelength of 530 nm and a 43 nm FWHM bandwidth, and is then detected by a CMOS camera. The CMOS camera can also be replaced by a photodiode for quantitative measurements or a scientific CCD camera (see pages 308 - 322) for higher-quality fluorescence images.

Schematic of a Typical Fluorescence Module

Objective

Tweezers Laser

Incident Radiation

Excitation Excitation Filter

Dichroic Mirrors Fluorescence (Emission) Emission Filter

CMOS Camera (or other detection system)

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Imaging Systems Bergamo Series Microscopes

Modular Optical Tweezers: Application Modules

Cerna Series Microscopes Essentials Kit Confocal Microscopes ThorImageLS Software Steering Module OCT Systems TDI Digital Microscopy Optical Tweezers

Galvo Mirror Control Module

Galvo Steering Module

Automated trap positioning capability can be added to the Modular Optical Tweezers system by integrating Thorlabs’ GVS002 2D galvo mirror (see pages 438 - 439), which replaces the turning mirror at the fiber input and is positioned in a plane conjugate to the back aperture of the objective. For the Keplerian configuration, rotations introduced by the galvo mirrors at a distance x from the first lens will be recreated at a distance y from the second lens according to equation y=

− (x−

)

( ⁄ lengths ) where f1 and f 2 are the focal of the two relay lenses. The magnitude of this rotation at location y is (f1 /f2 ) times the magnitude at location x.

GVS002

The example shown in the picture above is constructed with achromatic doublet 2D Galvo Mirror lenses. The first achromatic doublet is mounted into an adjustable lens tube and is positioned one focal length away from the center of the scanning mirrors. The multiplying factor to rotations created at the galvo mirrors position is 0.4. Drive voltages are applied to the galvo mirror controller boards via a DAQ card (not included), allowing the user to position the trap while the sample stage remains stationary. Due to the optical path length between the galvo mirror and the back aperture of the objective, only small angle adjustments are necessary, which means that the galvo mirror can operate at its maximum bandwidth of 1 kHz. By moving the beam back and forth between two positions with an appropriate dwell time at each position, it is possible to create two stable traps from a single laser beam. The images below display an example of two beads, 1 µm in diameter, that are simultaneously trapped by scanning the galvo mirrors at 200 Hz. The separation of the beads is about 6 µm. For information about this module or to request a quotation, please contact applications@thorlabs.com

Simultaneously Trapped 1 µm Beads

Time series images of Ø1 µm fused silica beads in a flow channel. The two beads circled in yellow are held by time-shared traps and remain stationary.

For more details regarding customization or modifications to the Modular Optical Tweezers, please contact the Advanced Applications Group at Applications@thorlabs.com 136

Raman Spectroscopy Application Module For customers who are considering adding Raman spectroscopy to their optical tweezers, contact applications@thorlabs.com for more information.

Imaging Systems Bergamo Series Microscopes

Optical Tweezers Sample Preparation Kit

Cerna Series Microscopes

Kit Contents Non-Drying Immersion Oil for Microscopy, Cargille Type B n Non-Functionalized Fused Silica Beads in Water, Ø1 µm, <0.5 g/mL n Mini Pipette with a 50 µL Volume n Two Plastic Slides with Built-In Channel, 400 µm Height, 100 µL Volume n 5 Microscope Glass Slides with Reaction Wells, Ø10 mm, 20 µm Deep n 100 Pieces of 18 mm x 18 mm Cover Glass, No. 1.5 Thickness n Dropper for Immersion Oil n

ITEM # OTKBTK

PRICE $ 145.00

Essentials Kit Confocal Microscopes ThorImageLS Software OCT Systems

OTKBTK

The OTKBTK Sample Preparation Kit is designed to allow users to quickly prepare a sample with Ø1 µm fused silica beads and test any optical tweezers system.

TDI Digital Microscopy Optical Tweezers

DESCRIPTION Sample Preparation Kit For OTM200 Optical Tweezers System and OTKB Optical Tweezers Kit

Microscopy Slide Holder Features n Accommodates

Glass Slides of Variable Width and a Length ≥44.0 mm (1.73") n Compatible Petri Dish Diameters from 37 to 41.4 mm (1.46" to 1.63") n Spring Clips Hold Sample Firmly in Place n Mounting Hole Compatibility • MAX Series Stages (See www.thorlabs.com) • Any Stage with 1/4"-20 (M6) Taps on 2" Centers n Dimensions: 101.6 mm x 68.6 mm x 12.7 mm (4" x 2.7" x 0.5") ITEM # MAX3SLH

PRICE $ 150.00

The MAX3SLH Microscopy Slide Holder MAX3SLH allows motion control stages to be used with petri dishes and glass slides. The stage/holder/slide assembly can then be integrated into custom-built microscopy setups like the optical tweezers. DESCRIPTION Microscopy Slide Holder

High-Numerical Aperture Objectives For applications that involve the optical trapping and manipulation of higher index particles such as polystyrene beads, we recommend a plan fluorite or semi-apochromatic objective that has a higher numerical aperture than the standard 1.25 NA objective included with the OTKB system. A higher NA objective will enable single particle spectroscopy, such as the trapping of a polystyrene probe bead, while simultaneously exciting and observing the fluorescence from the trapped bead.

N100X-PFO

ITEM # RMS100X-PFO

PRICE $ 2,644.00

DESCRIPTION 100X Olympus Plan Fluorite Oil Immersion Objective, 1.30 NA, 0.20 mm WD

N100X-PFO

$ 2,112.00

100X Nikon Plan Fluorite Oil immersion Objective, 1.3 NA, 0.16 mm WD

MOIL-30

45.00

RMS100X-PFO

Low Auto Fluorescence Immersion Oil, 30 mL

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