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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 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.

SUBMITTER: Cardoso Dos Santos M

PROVIDER: S-EPMC5034310 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature

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

Cardoso Dos Santos Marcelina M Déturche Régis R Vézy Cyrille C Jaffiol Rodolphe R

Biophysical journal 20160901 6

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 c ...[more]

PMID: 27653490

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