Several limitations restrict conventional Fluorescence recovery after photobleaching (FRAP) application. Examples are: 1) Low temporal resolution, which prohibits measurements of faster processes 2) FRAP evaluation schemes cannot include spatial constraints imposed by the cellular environment on protein mobility 3) FRAP ignores the sequential nature of the bleaching and image acquisition process. In order to overcome these limitations of conventional FRAP DKFZ researchers developed a novel FRAP based method called 3PEA (Pixel-wise Photobleaching Profile Evolution Analysis). The advantages of 3PEA are e.g.: accurate mobility measurements of fast, slow, and immobile proteins and fast determination of effective diffusion coefficients. The presented technology is thought to be suitable for use in all confocal laser scanning microscopes (no additional hardware is needed) and would allow automated high throughput FRAP experiments.
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DKFZ (German Cancer Research Center, Deutsches Krebsforschungszentrum)
Phone: +49-6221-42 2955
Dr. Ruth Herzog
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