Imaging with total internal reflection fluorescence microscopy for the cell biologist
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Imaging with total internal reflection fluorescence microscopy for the cell biologist.
Total internal reflection fluorescence (TIRF) microscopy can be used in a wide range of cell biological applications, and is particularly well suited to analysis of the localization and dynamics of molecules and events near the plasma membrane. The TIRF excitation field decreases exponentially with distance from the cover slip on which cells are grown. This means that fluorophores close to the ...
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Total internal reflection fluorescence microscopy (TIRFM) is an optical technique that allows imaging of a thin layer of the sample with a thickness of about 100-200 nm. It is used in science of cell biology to study cellular processes, especially near the membranes of living cells. This method is based on the total internal reflection phenomenon, where the evanescent wave is generated in the l...
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Total internal reflection fluorescence microscopy has been applied to image the final stage of constitutive exocytosis, which is the fusion of single post-Golgi carriers with the plasma membrane. The use of a membrane protein tagged with green fluorescent protein allowed the kinetics of fusion to be followed with a time resolution of 30 frames/s. Quantitative analysis allowed carriers undergoin...
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Total internal reflection fluorescence (TIRF) microscopy (TIRFM) is an elegant optical technique that provides for the excitation of fluorophores in an extremely thin axial region ("optical section"). The method is based on the principle that when excitation light is totally internally reflected in a transparent solid (e.g., coverglass) at its interface with liquid, an electromagnetic field, ca...
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The goal in fluorescence microscopy is to detect the signal of fluorescently labeled molecules with great sensitivity and minimal background noise. In epifluorescence microscopy, it is difficult to observe weak signals along the optical axis, owing to the overpowering signal from the out-of-focus particles. Confocal microscopy uses a small pinhole to produce thin optical sections ( 500 nm), but...
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ژورنال
عنوان ژورنال: Journal of Cell Science
سال: 2010
ISSN: 1477-9137,0021-9533
DOI: 10.1242/jcs.056218