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
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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|
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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