Topography of Cells Revealed by Variable-Angle Total Internal Reflection Fluorescence Microscopy
2016; Elsevier BV; Volume: 111; Issue: 6 Linguagem: Inglês
10.1016/j.bpj.2016.06.043
ISSN1542-0086
AutoresMarcelina Cardoso Dos Santos, Régis Deturche, Cyrille Vézy, Rodolphe Jaffiol,
Tópico(s)Cell Adhesion Molecules Research
ResumoWe 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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