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re SOLUTION 09 NANOTUBES AND SYNAPSES IN IMMUNE CELL COMMUNICATION Confocal Laser Scanning Microscopy
12 SEEING LIFE BEGIN Dynamic Imaging Sheds Light on Embryonic Development Processes
14 THE STEREOMICROSCOPE AS A 3D MEASUREMENT DEVICE
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Leica TCS SP2 SE Sensitivity Enhanced for Brilliant Images Sensitivity is the basis for high imaging quality. The new Leica spectral confocal TCS SP2 SE uses technologies to further improve image quality and significance of data. In addition to the well proven filter free AOBS® beam splitting and SP® tunable detection, the SE version is equipped with extra sensitive PMTs, and allows optional APD imaging at very high quantum efficiency and very low dark noise for brilliant images. The Super VISIR coating drastically improves transmission, especially in the infrared range for MP (multiphoton microscopy). The TCS SP2 SE offers better light sources as well: double power 405nm laser for DAPI, Hoechst, paGFP; and a high energy Ar-laser for FRAP experiments.
EDITORIAL
EDITORIAL Dear Reader Meeting the current and future needs of Leica customers was the driver for the reorganisation of Leica’s European Sales Divisions last year. In Life Science Research, Leica considered the changes to be a pre-requisite for commercial success as this could only be achieved by clearly understanding the needs of our customers and by providing leading edge products to satisfy those needs. Therefore, in July 2004, Leica formed a new European Research Sales Division totally dedicated to addressing Life Science Research in the area of Microscopy and High End Imaging Systems. The timing of the reorganisation was also influenced by the simultaneous introduction of a new generation of Microscopy Systems offering previously unavailable benefits to key areas of life sciences especially, but not only, in the area of Live Cell Imaging and Stem Cell Research. The development and introduction of this new generation of products, from Confocals to quantitative Widefield and advanced Fluorescence Systems, is ongoing with more ground breaking and unique products coming to market in the near future. In addition to product offerings, Leica also has a wealth of applications knowledge and high end imaging expertise, all of which may be of value to European scientists. As a vehicle to bring news of new product developments to the scientific community and to be able to share our knowledge and expertise we have decided to produce a European edition of our current Resolution magazine specifically for Life Science Research. This magazine will develop as we respond to reader feedback and will be published regularly from now on. I do hope you enjoy our first edition and it brings you some real value. John Buckley Director European Research Sales Division Leica Microsystems
Leica AF6000 Leica Microsystems introduces the new Leica AF6000 system for fluorescence imaging and live cell analysis. This extremely flexible system can be configured to all requirements for use in highly analytical applications, turning brilliant fluorescence images into significant data. Please see page 8 for further details.
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Welcome to the first publication of reSOLUTION – European Research Edition! This magazine is for you, dear customer. We have created this forum to provide you with product information and many of the upcoming European Leica Microsystems events over the next few months. Our goal is to publish this newsletter 2-3 times a year. Also in this first edition, we will present to you two researchers using Leica Microsystems products in their everyday work: First, Dr. Daniel Davis (Imperial College, London) who has kindly given us a recent article on Molecular Immunology. Secondly, you can read about developmental biology on zebrafish from the lab of Dr. Reinhard Koester (GSF Neuherberg, Germany). Finally, a contest where you can win great prizes, just for giving us your opinion, awaits you along with the calendar of events. So enjoy the newsletter, and I look forward to hearing from you.
NEW PRODUCTS
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APPLICATION REPORTS
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Leica DMI3000 B and DMI4000 B complement the Leica DMI6000 B The correct instrument for any biomedical application
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Confocal Laser Scanning Microscopy Nanotubes and Synapses in Immune Cell Communication
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Dynamic imaging sheds light on embryonic development processes: Seeing Life Begin
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New micromanipulation system by Leica and Eppendorf New system for fluorescence imaging and live cell analysis: AF6000
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The stereomicroscope as a 3D measurement device
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The world’s only interactive Leica 3D system for microscopy
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Leica MacroFluo™ – the new precision in fluorescence macroscopy
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EUROPEAN RESEARCH EVENTS
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CONTEST: Your opinion is valuable!
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IMPRINT
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CONTENTS
Paul Wismer, European Marketing Manager, Research, Leica Microsystems
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Range Expansion of Inverted Research Microscopes
Leica DMI3000 B and DMI4000 B Complement the Leica DMI6000 B Leica Microsystems has introduced a new line of inverted research microscopes for the discerning live cell researcher: The Leica DMI Series. Brilliance, convenience, integration, automation, and flexibility characterize the innovations.
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“Simply Microscopy“ is the central theme for Leica’s DMI microscopes. This phrase describes intelligent automation of the microscope, with the result that a researcher can concentrate fully on his or her experiment and not on microscope functions.
manual stand is also available for cost-conscious users. The Leica DMI3000 B is designed for customers that work exclusively with transmitted light methods. The new Leica DMI3000 B and DMI4000 B inverted microscopes not only exceed the latest technical standards, they also fulfil all ergonomic requirements to provide comfortable use during long periods of time at the microscope. In addition to live cell research, Leica DMI microscopes are ideal for routine examinations such as scanning cell or tissue cultures.
The Leica DMI research series, with its DMI3000 B, DMI4000 B, and DMI6000 B models, offers the right solution for all live cell applications. The DMI family includes fully automated versions, as well as ’coded’ systems that make it easier to select the correct components, and document your settings. A completely
Highly complex microscope processes are not only motorised in the DMI6000 B and DMI4000 B, but fully automated, so that users can concentrate fully on their research work. Leica’s intelligent automation still leaves room for individual requirements, however: manual intervention is possible at any time.
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Through improved optics and reduced background radiation, the Leica DMI4000 B and DMI6000 B achieve especially brilliant fluorescence. The fluorescence turret is equipped with up to six filter cubes that can be moved into position quickly, without vibration, at the touch of a button. The fluorescence axis also features an ultrafast, internal filter wheel (IFW) that supports the excitation of fluorochromes in less than 20 milliseconds. The Fluorescence Intensity Manager (FIM) reduces light stress for living cells and the bleaching of fluorochromes – the intensity of the excitation light can be reduced in a reproducible manner while the quality of the fluorescence is improved. The integrated, motorised Excitation Manager (ExMan) balances a variety of emission intensities to subsequently compensate for specimen artifacts. This innovative combination is truly unique. The microscopes are also designed to be completely light-tight, making it possible to mount even highly sensitive cameras on one of the four ports, without adversely affecting results. A versatile line of accessories, tailored to the microscope, create ideal conditions for live cell observation. These include incubators and heating/cooling devices, to provide ideal physiological conditions for cells at all times in regard to temperature, carbon dioxide, and oxygen ratios. Particularly for micromanipulation, Leica has designed especially narrow specimen stages that allow manipulators and other peripheral devices to be positioned close to the specimen. > VN
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Range Expansion of Inverted Research Microscopes
The Correct Instrument for Any Biomedical Application Leica Microsystems Inc, the expert in innovative, high-tech, precision optical instrument design, has expanded its line of DMI inverted research microscopes with the new Leica DMI3000 B and Leica DMI4000 B models. “Simply Microscopy” is the central theme for Leica’s DMI inverted microscope series, that focuses on simple operation through intelligent automation. The Leica DMI inverted research series, with its DMI3000 B, DMI4000 B and DMI6000 B models, offers a suitable solution for all live cell applications. Highly complex processes are not only motorised in the DMI6000 B and DMI4000 B, but fully automated, allowing users to concentrate completely on their research work. Leica’s intelligent automation still leaves room for individual requirements, however, and manual intervention is possible at any time.
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The Leica DMI family includes fully automated versions as well as ’coded’ systems that make it easier to select the correct components, and document your settings. A completely manual stand is also available for cost-conscious users. The Leica DMI3000 B is designed for customers that work exclusively with transmitted light methods. The new Leica DMI3000 B and DMI4000 B inverted microscopes not only exceed the latest technical standards, they also fulfil all ergonomic requirements to provide comfortable use during long periods of time at the microscope. The DMI series is designed for the study of live cells, micromanipulation, electrophysiology, and is also suitable for routine examinations such as scanning cell or tissue cultures. > VN
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New Micromanipulation System
New System by Leica and Eppendorf Leica Microsystems Inc. and Eppendorf present the second generation of their micromanipulation line. As the successor to the Leica AS TP Transgenic Platform, the Leica AM6000 is based on state-of-the-art technology – the Leica DMI6000 B inverted digital microscope. AM6000 users will benefit from all of the advantages of this fully automated research instrument. The comprehensive integration of microscope and micromanipulators in the new manipulation system offers a wide range of new possibilities."Simply Microscopy" is the central theme for the Leica DM line of research microscopes that focuses on simple operation through intelligent automation.
manipulation, but also the most important microscope functions. This integrated system accelerates work processes and enhances efficiency. In addition to all of the customary functions of NK2generation Eppendorf manipulators, the panels also feature focus handwheels and function buttons for brightness, objective selection, and the magnification changer. Objectives can be associated with contrast methods, stored with the magnification changer, and recalled as required. The Leica AM6000 is designed especially for transgenics and ICSI applications. > VN
The complete Leica AM6000 system is controlled from multifunction panels that cover not only micro-
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New AF6000 System
New System for Fluorescence Imaging and Live Cell Analysis From overlaying multi-channel images to z stacking and time lapse features, a wealth of solutions are included as standard for image documentation, quantification enhancement and analysis. Designed to completely harmonise microscope, camera and application, the Leica AF6000 is compatible with a wide range of Leica Microsystems fluorescence research microscopes and is well matched to both upright and inverted instruments. The AF6000 is based on a new software platform which is shared by all imaging products of Leica Microsystems. The simplicity of its highly intuitive interface with guided workflow ensures fast and effortless handling and guarantees brilliant results in a short time.
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Confocal Laser Scanning Microscopy
Nanotubes and Synapses in Immune Cell Communication Many of the key cell surface molecules involved in immune cell surveillance are now identified and one important new scientific frontier is to understand where and when each protein-protein interaction occurs to regulate cell functions. It is well established, for example, that white blood cells called Natural Killer (NK) cells recognize and kill tumor or virus-infected cells via different activating receptors or by detecting a loss of self-protein using inhibitory receptors but how cell-cell interactions over a few minutes leads to the decision whether or not a cell is diseased is far from understood. Thus, imaging has a major role to play in contemporary cell biology and one interesting theme to emerge is that immune cell communication is often accompanied by the segregation of proteins into micrometer-scale domains at an intercellular contact or immunological synapse.
Dr. Daniel M. Davis Molecular Immunology Imperial College London Department of Biological Sciences
Dan Davis obtained a BSc. in Physics from the University of Manchester in 1992 and a PhD. in Physics in 1995, working with David Birch at Strathclyde University. Then, as an Irvington Institute Research Fellow, he studied immunology with Jack Strominger at Harvard University. In December 1999 he returned to London to establish his own research group at Imperial College. In 2000, he won the Oxford University Press/Times Higher Education Supplement Science Writing Prize and, in 2002, was awarded a European Molecular Biology Organisation Young Investigator award. Dan is currently author or co-author of about 60 scientific papers in photophysics and immunology, including the first description of a Natural Killer cell immune synapse, studies of proteins transferring between immune cells, and observations of membrane nanotubes connecting immune cells.
Patterns of proteins that form at immune synapses result from intimate cooperation between several factors: Inter- and intramembrane protein binding, membrane mechanics, and cytoskeletal rearrangements all lead to a lateral sorting of proteins within their respective fluid membranes. Immune synapses extend several micrometers across intercellular contacts and, at least in some circumstances, specific patterns of proteins correlate with the ensuing intracellular signalling and cellular effector functions. Recently, for example, we have examined the membrane localisation of proteins that are up-regulated upon infection or tumour transformation and can activate human NK cells [Eleme et al., J. Exp. Med., 2004, 199:1005]. By using laser scanning confocal microscopy and electron microscopy we showed that these stress inducible ligands, MICA and ULBP1, 2, 3, are expressed in specific membrane microdomains (lipid rafts). The molecular mechanism by which these proteins are organised at the cell surface involves covalent attachment to specific lipids. It is known that viruses can prevent cell surface expression of the NK cell activating proteins to evade NK cell activation. Perhaps also some viruses or tumours evolved mechanisms to destabilize NK activation by
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The NK cell immunological synapse. Figure shows a B-cell line (221/Cw6-GFP) transfected to express GFP-tagged class I MHC protein (green) surrounded by three NK cells (YTS/KIR2DL1) transfected to express an inhibitory receptor. Lysotracker staining acidic organelles is shown in red and DNA, stained by Hoechst, is shown in blue. Clusters of GFP-tagged class I MHC protein at the intercellular contacts mark the immune synapses formed.
disrupting the cell surface organisation of these NK cell ligands. Our images also revealed an unexpected intercellular transfer of proteins during immune surveillance [Vanherberghen et al., Proc. Natl. Acad. Sci., 2004, 101:16873].
surface proteins following the formation of an immune synapse. We were able to make this discovery by using single photon-excitation resonance scanning confocal microscopy to image immune cell interactions in vitro very rapidly.
The functional consequences of the intercellular transfer of proteins during immune surveillance are largely unknown but perhaps, for example, tagging healthy cells could eliminate repeated scanning of the same cell.
The immune synapse is likely to have multiple functions with varying degrees of importance for different cell-cell interactions. In addition to controlling intracellular signals, the structure of the immune synapse may also play a role in directing the secretion of cytokines or lytic granules towards the target cell. To understand immune surveillance further, it will help to image several protein species simultaneously, with single-molecule resolution, second-scale time resolution, and with objective automated analysis of protein distribution. We also need to make real-time observations of synapse formation in living tissues, perhaps for example, using two-photon confocal microscopy. Our understanding of the spatio-temporal aspects of immune cell communication is only just beginning and the application of novel imaging technologies to this area of research is more than likely to throw up further unexpected observations in the near future.
In another recent article [Ă–nfelt et al., J Immunol, 2004, 173:1511], we built upon work published in an earlier neuroscience paper [Rustom et al., Science, 2004, 303:1007], by presenting evidence of a novel mechanism for intracellular communication in immunology. Nanotubular highways, or membrane nanotubes, were seen to connect multiple cells together and were readily formed between varieties of immune cell types. Nanotubes could be created upon disassembly of immune synapses, as cells move apart, and so nanotube networks could be assembled from transient synapses. It seems that nanotubes can transport membrane material along the surface of the tube as well as inside the tube. Therefore, nanotubes may provide a mechanism for the transport of cell
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Further Reading This article was published by: DM Davis, Trends in Immunology 2002, 23: 356 DM Davis & ML Dustin, Trends in Immunology 2004, 25: 323
Gemma L. Buckland, Bjorn Ă–nfelt & Daniel M. Davis Division of Molecular and Cellular Biology, Sir Alexander Fleming Building, Imperial College London, UK. E-mail: d.davis@imperial.ac.uk
Bebhinn Treanor and Bjorn Ă–nfelt use a Leica resonance-scanning confocal microscope to examine immune synapse assembly. Photo by David Bacon.
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Dynamic Imaging Sheds Light on Embryonic Development Processes
Seeing Life Begin Without dynamic imaging methods, capturing embryonic development processes would be unthinkable. To automate laborious timelapse experiments on living zebrafish embryos, the developmental biologists at the GSF (National Research Center for Environment and Health) in Neuherberg, Germany, tested a complete software-controlled system developed by Leica Microsystems. The motorised fluorescence stereomicroscope, fluorescence imaging software and digital camera work together in precision, easily handling even complex time-lapse and Z-stack experiments with quick filter change routines and deconvolution. Comprehensive all-around testing as well as specific tests to determine the precision of the interaction between hardware and software made it obvious that, in every respect, the Leica System enriches everyday scientific work in a zebrafish neuroimaging laboratory. It is a cost-efficient alternative to expensive instruments for intravital microscopic imaging.
The zebrafish – transparency in the world of biology Martin Distel of the Zebrafish Neuroimaging Group Reinhard Köster at the GSF Neuherberg uses the zebrafish as a model organism for intravital studies of genetic development. Native to India's Ganges river, this popular research subject is highly sought after in biology labs around the world. A look through the microscope reveals the secret of its popularity: the zebrafish embryo is transparent, making it possible to observe firsthand the development of organs, such as the eye, as it grows.
Leica’s complete system – put to the test To efficiently capture images of development processes, the software-controlled system from Leica Microsystems was tested in various experiments on living zebrafish embryos. The FW4000 software was able to reliably automate all time-lapse experiments in bright-field and fluorescence modes and could handle the imaging of rapidly occurring processes, such as the various phases of the heartbeat (Fig. 1). Even complex time-lapse experiments with three fluorescence filters functioned perfectly (Fig. 2). UVconvertible pigments in cells of a zebrafish embryo were excited in succession with GFP3, DAPI and Texas Red. The motorised filter change was fast and so free of vibration that high-quality images could be obtained without any pixel shift.
The supreme discipline: complex experiments with filter change, Z-stack and deconvolution
Fig. 1: Time-lapse images of a zebrafish embryo with injected contrast medium
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The motorised focusing drive on the Leica MZ16 FA allows automated imaging in various planes along the Z-axis. In the test, a Z-stack in 15 planes was taken of
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Fig. 2: Complex time-lapse experiment with 3 fluorescence filters (GFP3, DAPI, Texas Red). Automated contrast medium conversion. Embryo before, during and after irradiation
MZ16 FA fluorescence stereomicroscope, can open up new paths in development research. In spite of its complexity, the software is quite easy to learn and use. The quality of the images achieved and the performance of the complete Leica system convinced the researchers at GSF Neuherberg. > IH
Fig. 3: Zebrafish embryo with multicoloured fluorescence-marked nuclei and cellular membranes; level 8 of the Z-stack a) GFP3 channel with fluorescence-marked green cell nuclei b) Texas Red channel with fluorescencemarked red cellular membranes c) Composite of a and b before deconvolution d) After deconvolution. Yellow areas have disappeared almost completely, i.e. the fluorescence signals are correctly assigned to their focal plane.
a doubly marked zebrafish embryo (Fig. 3). With the FW4000 software, the focal displacements and the change routine for the two fluorescence filters functioned reliably and without vibration. Then, the images from the individual channels (GFP3 and Texas Red) for each Z-plane were composed into a single image using the FW4000 software (Fig. 3c). This can produce a blurry image, as is the case here, because regular images taken with a stereomicroscope contain a lot of information from areas outside of the focal plane. Also, the overlapping “red and green� information creates a yellow colour that misleadingly indicates a colocalisation of nuclei and cellular membranes. The Leica Deblur imaging software provides the solution to this problem with its deconvolution function, which assigns non-focused light signals to their original focal plane. The deconvolution worked very well. Not only was the composite sharper (Fig. 3d), the subcellular structures also appeared in convincingly high resolution, and the yellow areas disappeared almost completely.
Experiments were convincing The software package from Leica Microsystems is very promising indeed for automating time-lapse and Z-stack experiments, and in combination with the
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The Stereomicroscope as a 3D Measurement Device Introduction Traditionally microscopists have used their stereomicroscopes for stereoscopic viewing and 2D imaging. There are countless uses for the stereomicroscope to examine the surfaces of specimen, but the challenge has always been the microscope’s inability to measure (and even image) in 3D. In stereomicroscopy the depth of focus is large, however no direct 3D visualisation and measurement are possible throughout this extended depth of focus. Consequently, when quantitative 3D surface analysis is desired, an alternative device is needed to supplement the stereomicroscope, or other solutions have to be applied. This simple limitation has posed an extreme challenge to scientists and their budgets of time, money‌and results. In this report a new 3D analysis capability in stereomicroscopy from Leica Microsystems is presented. Conventional stereomicroscopes can be transformed into true 3D measurement devices with full analytical capability to generate profile, roughness, area and even volumetric measurements. The Leica 3D system is useful in all fields of science and technology in which representation of spatial depth information means having more, and more valuable, information.
constructions. The integrated digital Leica IC 3D stereo camera creates a perfect high-resolution stereopair without the need for complex adjustments (Figure 1). Conventional stereoscopic approaches require accurate optical setups in order to get acceptable results. Developments by Leica Microsystems greatly reduce the requirements of that because an automatic offset calculation and geometric correction are performed. This calculation produces highly accurate and robust 3D surface reconstructions. The relative height accuracy is better than 3% and a fully automatic calculation can be performed in less than 100 seconds. The software can be applied to any stereomicroscope of the Leica Microsystems MSeries. The version for the Leica MZ16 A automated stereomicroscope controls not only the Leica IC 3D stereo camera, but also the motorised zoom and updates the data on the display. In addition, the autofocus module allows precision control of the motorised focus. These settings can be saved and retrieved as required. This makes work ergonomic and efficient, particularly during precise, repetitive experiments.
3D Stereo Reconstruction Currently no real 3D analysis is commercially available. This limitation is circumvented with the standalone software package Stereo Explorer from Leica Microsystems. This modular software automatically calculates a 3D data record from two partial images for reconstruction, documentation, analysis and quantitative measurement of three-dimensional surfaces. The realistic image, which appears in relief, makes it easier for the user to identify complex surfaces, greatly improves education and training, and enables better diagnoses in technical, biological and forensic fields. Stereopairs with excellent-quality, true-to-detail images are an essential requirement for precise 3D re-
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Fig. 1: The optical concept of 3D image capture: The Leica IC 3D digital stereo camera is integrated between the binocular tube and optics carriers of a Leica M Series stereomicroscope and takes a high-resolution image of each optical beam path.
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Fig. 2:Profile of a geological specimen
Quantitative measurement Once the surface information of the object is determined, the 3D data can be visualised and analysed. All analysis routines are designed in a modular, intuitive manner, and are supported with full windows reporting conventions. The possible analytical capabilities comprise numerous calculations where the description of such would go beyond the limits of this publication. The profile analysis module permits the extraction of height profile along user defined paths and can be used to calculate relative height measurements as shown in figure 2. All established roughness, profile and waviness
measurements can be performed as well as various statistical calculations. The area analysis module similarly calculates parameters such as the ratio of true-to-projected area, also termed the bearing area curve. The volume analysis module as shown in figure 3 enables direct volumetric measurements relative to freely definable polygon lines.
Applications The technology presented in this article can be applied in any field of study where the surface structure of a specimen is important. This may include documentation of the roughness of microscopic specimens for affinity calculations, the analysis of fractured surfaces from mechanical parts, or the volumes of im-
Fig. 3: Volume measurement in fracture analysis. A 3D model visualises the volume of interest.
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Fig. 4 3D model of a solder paste with axes of coordinates visualised with Leica Stereo Explorer
pressions and uprisings of formed surfaces. The special advantage of this innovative technology is the ability to establish, for the first time in microscopy, a direct link between the microscope image and the 3D measurement capability. There is no longer the need to struggle to find the area of interest in another device, the image and 3D surface data coexist. Some of the many applications might be: • • • • • • • •
Fig. 5 Height-encoded phantom colour view of a crystal
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Fracture analysis Materials development Drug discovery and production Semiconductor Life sciences research MEMS Entomology Geology
Summary Advances in image processing and computer technology permit measurement of 3D surface parameters directly in stereomicroscopic images. Profile, area and even volumetric calculations can be performed in reconstructed 3D images. Therefore the specimen does not need to be examined with other 3D measurement devices; rather all analysis can be performed with existing stereomicroscopes retrofitted with the new Leica Microsystems technology. This reduces investment costs while dramatically improving lab performance. All the measured values, the 3D models and other parameters can easily be exported to standard files, integrated and further processed in reports. This new technology for stereomicroscopy by Leica Microsystems will become a laboratory standard for advanced research. > DG
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The World’s Only Interactive Leica 3D System For Microscopy A capable and innovative partner in all aspects of 3D visualisation, Leica Microsystems has developed the world’s only 3D system for Leica M stereomicroscopes which allows real 3D visualization without the need for uncomfortable special glasses. The Leica 3D system is a professional, fully automatic solution for reconstruction, viewing, documentation, reporting, quantitative analysis and 3D measurement of three-dimensional surfaces and useful in all fields of science and technology. For visualising and analysing specimen surfaces with an accuracy that, until recently, was nearly unthinkable, the innovative Leica 3D system now makes the process easier, more effective and more precise. In addition to imaging of surfaces, contact-free quantification of surface parameters such as height differences, roughness, surface areas or volumes is also possible.
The three-dimensional image is created from a stereopair taken by the Leica IC 3D digital stereo camera. The intuitive and user-friendly software controls the Leica IC 3D camera and offers a variety of functions for optimally adjusting the imaging quality of the system to obtain a perfect 3D image. The Leica StereoExplorer software package automatically determines which pixels in the two slightly different images belong together, and calculates the topography of the specimen as a surface model, taking into consideration the parameters of angle and magnification level. The 3D data record thus created serves as the basis for surface and volume analyses. The optional the high-resolution Leica ASD Auto-Stereoscopic Display System offers real time display of specimens in all three dimensions. Objects appear as concrete and vivid as in real life – no eyewear or other accessories are necessary. The depth of field and colour reproduction correspond to the view through the eyepieces of a stereomicroscope. > IH
Leica MacroFluo™ – The New Precision in Fluorescence Macroscopy Leica Microsystems presents the world's only macro documentation systems for fluorescence methods, which have been developed in cooperation with the Imaging Centre of IGBMC. Leica MacroFluo™ systems with apochromatic manual or motorised 6.3:1 or 16:1 zooms are the right choice for observing and digitally recording the effects of genetic defects in living transgenic models the size of a whole mouse (zebrafish, Xenopus) with the highest precision and resolution in an intensely fluorescent field. The MacroFluo™ concept combines the advantages of macroscopy – parallax-free imaging, generous fields of view and long working distances – with fluorescence methods, including multicolour fluorescence, and high resolution. Unlike stereomicroscopy with two convergent optical paths, it guarantees an absolute maximum in precision when examining whole, living models, for measurements, and for digital image processing and analysis. That applies especially to processing with multifocus, overlay and deconvolution programs. Leica MacroFluo™ systems deliver highest macroscopic resolution for the experimental documentation of large objects: 340 Lp/mm with the 6.3:1 zoom and
336 Lp/mm with the 16:1 zoom (values with planapochromatic 1x objective and HC Plan wide-field eyepieces 10x/25). With the planapochromatic 5x HR (high resolution) microscope objective, the wealth of visual information delivered by the MacroFluo™ systems reaches the levels of traditional microscopy. Magnifications of up to 225x (6:1 zoom) or 575x (16:1 zoom) can be achieved with a resolution of 1500 Lp/mm. The bright, homogeneous fluorescence field automatically adjusts to match the field of view when zooming, guaranteeing precise illumination and maximum light utilisation at any zoom position. The 5-position rapid filter changer supports the use of 26 different fluorescence filters for a wide range of fluorescence techniques. The depth of field can be adjusted with the integrated iris diaphragm. Leica Microsystems offers a comprehensive range of innovative accessories for custom-tailored solutions to meet any requirement or solve any problem. Highresolution FireWire cameras for fluorescence and software (FW4000, IM1000 image overlay and multifocus) are available especially for documentation and image analysis. > IH
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EVENTS
European Research Events Here is just a sampling of some of the events Leica Microsystems will participate in or organise in the 2nd half of 2005. For further information, please visit our website: www.leica-microsystems.com (click on events on the left) Exhibits: Zebrafish – Genetics and Developmental Biology July 13–16 Dresden, GERMANY
Société Française de Biologie du developpement October 1–4 Obernai, FRANCE
BioScience 2005 July 17–21 Glasgow, UK
Scanlab 2005 October 11–13 Stockholm, SWEDEN
ELSO September 3–6 Dresden, GERMANY
BioTechnica 2005 October 18–20 Hannover, GERMANY
MAF9 September 4–7 Lisbon, PORTUGAL
“Simply Brilliant” Workshops:
Transgenic Tech meeting September 11–13 Barcelona, SPAIN Congress Nazionale di Neuroscienze 2005 October 1–4 Ischia, ITALY
GERMANY CW 28: Kiel, Biocentre CW 36: Berlin CW 37: Hamburg CW 39: Tübingen CW 45: Berlin, Charité, Mitte (Laser Microdissection Workshop) For other countries please contact your local Leica organisation.
CONTEST Win a pair of Leica Binoculars, Leica Wristwatch or other great prizes! Dear Customer: Please give us your comments on this first European Research Edition of reSOLUTION magazine. Send us your complete name and address of the institute where you work, along with your comments by going to the following link: www.leica-microsystems.com/EU-Research by August 31, 2005. Winners will be drawn on or around September 1, 2005 from all completed entries. 18 reSOLUTION
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PAT H F I N D E R S O F T WA R E
New Cell Recognition Software
Pathfinder Edition Together with the highly automated laser microdissection system AS LMD, Leica now offers microdissection with completely automated and integrated cell recognition. Collecting thousands of cells is now possible in just a few steps. The fully integrated software has a very powerful teaching mode to define the required target cells by a few simple mouse clicks.
Top quality downstream kits from Qiagen – for reliable results The Qiagen integrated solution for RT analysis gives LMD users ultra high sensitivity and specificity for downstream analysis in RNA and DNA research. No optimisation is required. Optimised protocols are directly available together with professional application support from Qiagen. The kits are specially designed for Leica Laser Microdissection.
Special 63x XT objective for Laser Microdissection Dissecting single cells in the range of 5–20 µm in diameter requires special optics. With our 63x XT HCX PL FLUOTAR objective, these requirements are met with brilliant image quality. The large working distance of this objective allows easy and safe microdissection when working with groups or individual cells.
High quality service support for even longer than usual Leica’s excellent worldwide service helps you to keep your equipment running and up to date. With cutting edge technologies like remote service via internet, support can be provided quickly and easily. The modular system can easily be serviced by part exchange. The extended warranty will give you this performance for one additional year.
Imprint reSOLUTION is the magazine for Leica Microsystems customers and business friends Publisher Leica Microsystems AG, Wetzlar (Germany) Editors Paul Wismer Ralf Niggemann Editorial address Leica Mikrosysteme Vertrieb GmbH Lilienthalstraße 39 – 45 DE-64625 Bensheim Tel.: +49 (0)6251-136-210 Fax +49 (0)6251-136-155 www.leica-microsystems.com E-Mail: Paul.Wismer@leica-microsystems.de Layout & Production Central Marketing Uwe Neumann Leica Microsystems AG Ernst-Leitz-Strasse 17 – 37 DE-35578 Wetzlar Published twice or three times a year Cover Picture Leica Microsystems AG Fluorescence Applications Contributing editors John Buckley > JB Verena Nickl > VN Christine Ludwig > CL Ingrid Haack > IH Daniel Davis > DD Daniel Göggel > DG
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reSolution Newsletter2_01_05
14.07.2005
9:12 Uhr
Seite 20
Leica & Science! Famous scientists work with instruments from Leica Microsystems Leading scientists who use instruments from Leica Microsystems value their high quality, technical perfection, technological standards, service and large selection. When designing a new product, Leica Microsystems always puts the user first and offers the best possible solution for each application and every budget: from routine microscope to high end systems for life science and industry.
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