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Nanoscale topography of cells revealed by variable-angle Total Internal Reflection Fluorescence Microscopy (vaTIRFM)

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Abstract

We propose an improved version of variable-angle total internal reflection fluorescence microscopy (vaTIRFM) adapted to modern TIRF setup. This technique involves the recording of a stack of TIRF images, by gradually increasing the incident angle of the light beam on the sample. A comprehensive theory was developed to extract the membrane/substrate separation distance from fluorescently labeled cell membranes. A straightforward image processing was then established to compute the topography of cells with a nanometric axial resolution, typically 10–20 nm. To highlight the new opportunities offered by vaTIRFM to quantify adhesion process of motile cells, adhesion of MDA-MB-231 cancer cells on glass substrate coated with fibronectin was examined.
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Dates and versions

hal-02460110 , version 1 (29-01-2020)

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  • HAL Id : hal-02460110 , version 1

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Dalia El Arawi, Marcelina Cardoso dos Santos, Régis Deturche, Cyrille Vézy, Rodolphe Jaffiol. Nanoscale topography of cells revealed by variable-angle Total Internal Reflection Fluorescence Microscopy (vaTIRFM). Focus on microscopy 2017, Apr 2017, Bordeaux, France. ⟨hal-02460110⟩

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