Authors
Marcelina Cardoso Dos Santos, Régis Déturche, Cyrille Vézy, Rodolphe Jaffiol,
Title
Topography of cells revealed by variable-angle Total Internal Reflection Fluorescence Microscopy
In
Biophysical Journal
Volume
111
Issue
6
Pages
1316–1327
Publisher
Cell Press
Year
2016
Indexed by
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.
Affiliations
Offprint