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Photobleaching methods in cell biology

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Abstract

Techniques have emerged in the past decade to measure the translational mobility of fluorescence- labeled molecules within microscopic specimens including the membranes and cytoplasm of single living cells. The methods are based on photo- bleaching discrete regions of the samples bearing fluorescent molecules, thereby destroying their emission, and the subsequent observation of the recovery of fluorescence due to the diffusion or flow of unbleached fluorophores from the surrounding region into the previously bleached area. The time it takes for recovery to occur is directly related to the diffusion coefficient or flow velocity of the labeled molecule. At this time, no other methods give comparable information about the translational mobility of molecules in defined regions of individual living cells and their organelles.

© 1985 Optical Society of America

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