Fluorescence Microscopy: Applications of Fluorescence Recovery after Photobleaching to protein transport in living cells.

By:

Shardrack Asare

Abstract

Fluorescence recovery after photobleaching (FRAP) is a non-invasive analytical technique that utilizes a confocal microscope and green fluorescent protein to study the rate of diffusion and dynamics through tissue or cells. In most FRAP experiments, proteins tagged with green fluorescent protein (GFP) is illuminated with a laser source, causing a bleach of the GFP but not the protein’s function, the rate of protein diffusion can then be determined by photobleaching a specific area of the cytoplasm and measuring the rate of fluorescence recovery. In this talk, we will discuss the use of FRAP to measure the mobility of GFP-tagged Sup35p in its non-prion form in [psi^-] and its prion form in [PSI⁺] cells. In conclusion, FRAP has successfully been used to measure aggregation state of [PSI⁺], and this has giving a better understanding of the mechanism of prion curing by guanidine hydrochloride (Gdn).

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