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.
journal_name
Biophys Jjournal_title
Biophysical journalauthors
Cardoso Dos Santos M,Déturche R,Vézy C,Jaffiol Rdoi
10.1016/j.bpj.2016.06.043subject
Has Abstractpub_date
2016-09-20 00:00:00pages
1316-1327issue
6eissn
0006-3495issn
1542-0086pii
S0006-3495(16)30613-0journal_volume
111pub_type
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