title: 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.
authors.affiliation: Laboratoire de Nanotechnologie et d'Instrumentation Optique, Institut Charles Delaunay - UMR 6281 Centre National de la Recherche Scientifique, Université de Technologie de Troyes, Troyes, France.
authors.forename: Marcelina
authors.initials: M
authors.lastname: Cardoso Dos Santos
authors.affiliation: Laboratoire de Nanotechnologie et d'Instrumentation Optique, Institut Charles Delaunay - UMR 6281 Centre National de la Recherche Scientifique, Université de Technologie de Troyes, Troyes, France.
authors.forename: Régis
authors.initials: R
authors.lastname: Déturche
authors.affiliation: Laboratoire de Nanotechnologie et d'Instrumentation Optique, Institut Charles Delaunay - UMR 6281 Centre National de la Recherche Scientifique, Université de Technologie de Troyes, Troyes, France.
authors.forename: Cyrille
authors.initials: C
authors.lastname: Vézy
authors.affiliation: Laboratoire de Nanotechnologie et d'Instrumentation Optique, Institut Charles Delaunay - UMR 6281 Centre National de la Recherche Scientifique, Université de Technologie de Troyes, Troyes, France. Electronic address: rodolphe.jaffiol@utt.fr.
authors.forename: Rodolphe
authors.initials: R
authors.lastname: Jaffiol
mesh_terms: D000465:Algorithms; D002138:Calibration; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002462:Cell Membrane; D048430:Cell Shape; D004574:Electromagnetic Fields; D004867:Equipment Design; D005353:Fibronectins; D005453:Fluorescence; D005898:Glass; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008962:Models, Theoretical
chemical_list: D005353:Fibronectins