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Journal Articles Biophysical Journal Year : 2016

Topography of Cells Revealed by Variable-Angle Total Internal Reflection Fluorescence Microscopy

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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 sep- aration 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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hal-02530478 , version 1 (03-04-2020)

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Marcelina Cardoso dos Santos, Régis Deturche, Cyrille Vézy, Rodolphe Jaffiol. Topography of Cells Revealed by Variable-Angle Total Internal Reflection Fluorescence Microscopy. Biophysical Journal, 2016, 111 (6), pp.1316-1327. ⟨10.1016/j.bpj.2016.06.043⟩. ⟨hal-02530478⟩

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