Total internal reflection fluorescence microscopy: application to substrate-supported planar membranes.

N L Thompson; K H Pearce; H V Hsieh

doi:10.1007/bf00213560

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Total internal reflection fluorescence microscopy: application to substrate-supported planar membranes.

N L Thompson·K H Pearce·H V Hsieh

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Abstract

The use of total internal reflection illumination in fluorescence microscopy (TIRFM) is reviewed with emphasis on application to fluorescent macromolecules that specifically and reversibly bind to planar model membranes supported on glass or quartz substrates. Several methods for characterizing macromolecular motion and organization are discussed: the measurement of equilibrium binding curves to obtain values for equilibrium binding constants; the measurement of fluorescence photobleaching recovery curves to obtain values of kinetic rate constants and surface diffusion coefficients; and the measurement of fluorescence intensities as a function of the evanescent field polarization to characterize orientational order. Applications to cell-substrate contact regions are summarized and future directions of TIRFM are outlined.

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Chemical Phenomena Chemistry, Physical Fluorescence Membranes, Artificial Microscopy, Fluorescence Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.Review

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N L Thompson, K H Pearce, H V Hsieh. 1993. Total internal reflection fluorescence microscopy: application to substrate-supported planar membranes.. https://doi.org/10.1007/bf00213560

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