Photoswitching-free FRAP analysis with a genetically encoded fluorescent tag.

ABSTRACT: Fluorescence recovery after photobleaching (FRAP) is a widely used imaging technique for measuring protein dynamics in live cells that has provided many important biological insights. Although FRAP presumes that the conversion of a fluorophore from a bright to a dark state is irreversible, GFP as well as other genetically encoded fluorescent proteins now in common use can also exhibit a reversible conversion known as photoswitching. Various studies have shown how photoswitching can cause at least four different artifacts in FRAP, leading to false conclusions about various biological phenomena, including the erroneous identification of anomalous diffusion or the overestimation of the freely diffusible fraction of a cellular protein. Unfortunately, identifying and then correcting these artif

SUBMITTER: Morisaki T 

PROVIDER: S-EPMC4169462 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature