The physical basis of total internal reflection fluorescence (TIRF) microscopy and its cellular applications. 2015

Natalie S Poulter, and William T E Pitkeathly, and Philip J Smith, and Joshua Z Rappoport
Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.

Total internal reflection fluorescence (TIRF) microscopy has gained popularity in recent years among cell biologists due to its ability to clearly visualize events that occur at the adherent plasma membrane of cells. TIRF microscopy systems are now commercially available from nearly all microscope suppliers. This review aims to give the reader an introduction to the physical basis of TIRF and considerations that need to be made when purchasing a commercial system. We explain how TIRF can be combined with other microscopy modalities and describe how to use TIRF to study processes such as endocytosis, exocytosis, and focal adhesion dynamics. Finally, we provide a step-by-step guide to imaging and analyzing focal adhesion dynamics in a migrating cell using TIRF microscopy.

UI MeSH Term Description Entries
D008856 Microscopy, Fluorescence Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye. Fluorescence Microscopy,Immunofluorescence Microscopy,Microscopy, Immunofluorescence,Fluorescence Microscopies,Immunofluorescence Microscopies,Microscopies, Fluorescence,Microscopies, Immunofluorescence
D002462 Cell Membrane The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells. Plasma Membrane,Cytoplasmic Membrane,Cell Membranes,Cytoplasmic Membranes,Membrane, Cell,Membrane, Cytoplasmic,Membrane, Plasma,Membranes, Cell,Membranes, Cytoplasmic,Membranes, Plasma,Plasma Membranes
D003584 Cytological Techniques Methods used to study CELLS. Cytologic Technics,Cytological Technic,Cytological Technics,Cytological Technique,Technic, Cytological,Technics, Cytological,Technique, Cytological,Techniques, Cytological,Cytologic Technic,Technic, Cytologic,Technics, Cytologic
D003585 Cell Biology The study of the structure, behavior, growth, reproduction, and pathology of cells; and the function and chemistry of cellular components. Cellular Biology,Biologies, Cell,Biologies, Cellular,Biology, Cell,Biology, Cellular,Cell Biologies,Cellular Biologies
D022001 Focal Adhesions An anchoring junction of the cell to a non-cellular substrate. It is composed of a specialized area of the plasma membrane where bundles of the ACTIN CYTOSKELETON terminate and attach to the transmembrane linkers, INTEGRINS, which in turn attach through their extracellular domains to EXTRACELLULAR MATRIX PROTEINS. Adhesion Plaques,Cell-Matrix Adherens Junctions,Focal Contacts,Adherens Junction, Cell-Matrix,Adherens Junctions, Cell-Matrix,Adhesion Plaque,Adhesion, Focal,Adhesions, Focal,Cell Matrix Adherens Junctions,Cell-Matrix Adherens Junction,Contact, Focal,Contacts, Focal,Focal Adhesion,Focal Contact,Junction, Cell-Matrix Adherens,Junctions, Cell-Matrix Adherens,Plaque, Adhesion,Plaques, Adhesion

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