Fluoresence recovery afer photobleaching

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Fluoresence recovery afer photobleaching technoque was developed around 1976 in the lab of Watt W.Webb when they were interested in the lateral mobility of the acetylcholine receptor(AChR). The introduction of green fluorescent protein (GFP), and the development of commertially available confocal microscope based photobleaching methods have lead to the revival of fluorescence technique.Using this technique, a region of interest is selectively photobleached with a high-intensity laser, and the recovery that occurs as fluorescent molecules move into the bleached region is monitored over time with low-intensity laser light. Complementary to the photobleaching techniques discussed earlier, the continuity of a cell compartment can be monitored using a technique called Fluorescence Loss In Photobleaching (FLIP).