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- W2081123589 abstract "Many cellular mechanosensory functions rely on molecular interactions related to cellular force exertion. Despite the rich biochemical knowledge in this field, the current mechanistic picture - that can be enhanced by high-resolution optical microscopy techniques - is still in its infancy. Here, we describe a simple yet universal approach to combine high-resolution optical microscopy techniques with traction force measurements. Etching 50 μm high spacers next to a micropillar array and inverting the array onto a thin coverslip before imaging enables the use of a short working distance objective. In our setup we use a spinning-disk confocal on an inverted microscope with a 100X, NA 1.4 objective. Our deflection detection resolution per micropillar is below 30 nm, corresponding to a force resolution of 500 pN. We validate our technique by imaging fixed and live mouse fibroblasts with dye-conjugated fibronectin micro-contact printed on the micropillars and immunostaining for paxillin and actin. We observe forces of up to 15 nN on individual pillars using a precise force-deflection relationship calibrated using scanning electron microscopy, tensile testing and finite element analysis. We quantified focal adhesion growth with increasing force of 4 ± 1 nN/μm2 and co-orientation of focal adhesion elongation and force direction within 2 ± 24 degrees. Furthermore, the cells remain viable during overnight live-cell imaging and stay adhered to the substrate. Our novel approach opens up the combined application of traction force measurements and super-resolution optical microscopy techniques." @default.
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- W2081123589 date "2013-01-01" @default.
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- W2081123589 title "High-Resolution Fluorescence Measurements Correlated to Cellular Traction Forces" @default.
- W2081123589 doi "https://doi.org/10.1016/j.bpj.2012.11.1090" @default.
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