Patlisci2

Physics UNIBAS

Nano-Tera Annual Meeting 2015 Progress Report PatLiSci II P. Oertle, 04.05.2015 PATLiSci 2

PATLiSci II Probe Array Technology for Life Science E. Meyer (PI) Uni Basel Cantilever sensors R. Lim M. Loparic Uni Basel E-Mapping

Industrial partner:

U. Matter Nanosurf AG

N. de Rooij EPFL-IMT, MEMS design & fab

H. Heinzelmann CSEM Probe array technologies End-Users:

K. Glatz University Hospital Basel Pathology

P. B. Henrich University Hospital Basel Ophthalmology

R. Zanetti University Hospital Basel Oncology

PATLiSci 2

Outline/Workflow

High-Tech Biomarker Sensing

Cutting Edge Mechanical Markers

RAPID CANCER DETECTIO N

Multiplexe d Atomic Force Microscopy

PATLiSci 2

Cancer Stiffness is a Diagnostic Marker

Tumor invasion and metastasis depends on the mechanical properties of cancer cells.

Fuster & Esko Nature Reviews Cancer 2005 Butcher et al Nature Reviews Cancer 2009; Fritsch et al Nature Physics 2010; Wirtz et al Nature Reviews Cancer 2011

Feeling the Force of Cancer

Harvest Biopsy

Measure

Reconstruct

1 map = 1024 force measurements

Nanomechanical signature of cancer Cancer "aggressiveness" peak

Plodinec et al., Nature Nanotechnology (2012)

Cantilever Array for Cancer Detection Silicon nitride probe 500 μm x 100μm Pitch 250 μm ~0.05 N/m x8 cantilevers

 Cantilever array for fast, parallelized detection of cancer  Optimal probe specifications for force spectroscopy and cantilever sensing

An in vitro epithelium that bears the mechanobiological hallmarks of living tissue The Inner Limiting Membrane (ILM) ZO-1

Laminin

DAPI/Laminin-ILM

Collagen IV ARTIDIS-AFM Stiffness Mapping

E-cad

Vinculin +Actin 5 μm α6 β4 Laminin

Marija Plodinec & Philipp Oertle

PATLiSci Arrays Can Measure an In Vitro Epithelia Comparison of PATLiSci Arrays (rectangular CL) to single CL (DNP-s10, Bruker triangular)

Both cantilevers give the same results!

Detection of BRAF Mutation (melanoma) and HER2 (breast cancer) in Biopsy RNA

Punch Biopsy

Cantilever Sensors total RNA

Measurement Baseli ne

PATLiSci 2

Physics UNIBAS

Detection of BRAF Mutation development of lifetime risk for melanoma in the last 50 years

rate of incidence

0.010

 BRAF V600 mutation is found in 50 % of patients

0.008 0.006

 Survival benefit increases significantly with inhibitor treatment

0.004 0.002 0.000

1960

2006

year M. Volkenandt: Maligne Melanome. In: Dermatologie und Venerologie.

O. Braun-Falco u. a. (Ed.), Verlag Springer, 2005, S. 1313–1324

Patient sample (mutant & wild-type) sample is injected

mtCl bends wtCl also bends refCl bends a little

Physics UNIBAS

Analysis of BRAF Biopsies in a double-blind study (BRAF mutation) Evaluation of wt sample

Number

Total RNA analysis

Evaluation of 4b (mt)

Amount of tot. RNA in Âľg

Lab Nr.

RNA conc. in ng/ul Histology Evaluation

01.02 Form.

9.4

1b

188

mt

mt

01.03 Frozen

48.45

2a

969

wt

wt

01.04 Form.

78.15

2b

1563

wt

wt

01.05 Frozen

11.7

3a

234

wt

wt

01.06 Form.

417

3b

8340

wt

wt

01.07 Form.

626.95

4b

12539

mt

mt

F. Huber et al 2015 PATLiSci II K. Glatz (University Hospital Basel)

6

6

Physics UNIBAS

Quantification of HER2 (over-)expression in breast tumor tissue

ď ą HER2 overexpression is found in 20 % of breast cancer patients ď ą Can be treated with Herceptin

RNA isolated from an invasive ductal carcinoma (malignant) and a fibroadenoma (benign)

F. Huber, M. Plodinec et al. 2015

Physics UNIBAS

Demonstrator Photodiode VCSL Positioning

Sample holder

VCSL Driving

Cantilever

VCSL Packaging LMIS4-CSEM | Master Project Defense Jonathan Cottet | 30.03.2014|

Page 14

Technology Transfer to Nanosurf FlexAFM

Physics UNIBAS

Conclusions PATLiSci 2

• Detection of breast cancer cells based on elasticity

RAPID CANCER DETECTION

• Sensing of cancer in patient biopsies achieved • Cantilever array delivered and tested

Integrate

• Demonstrator functional Parallelize

• Technology integration into NanoSurf FlexAFM designed

Sense Cancer Feel Cancer

PATLiSci 2