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

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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https://hal-utt.archives-ouvertes.fr/hal-02530478
Contributor : Jean-Baptiste Vu Van <>
Submitted on : Friday, April 3, 2020 - 8:02:44 AM
Last modification on : Saturday, April 4, 2020 - 1:44:40 AM

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

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