FRET Imaging Microscopy

摘要

Fluorescence Resonance Energy Transfer (FRET) Microscopy has been finding substantial utility in the measurement of a number of biological processes. Most microscopic techniques that have been developed to monitor FRET measure changes in the donor and acceptor emission or fluorescent lifetime of the donor. These include measurements of sensitized emission, acceptor photobleaching and fluorescent lifetime imaging (FLI). However, which of these approaches is the best for a given experimental situation and for use with multiphoton microscopy is not clear. Using mutant GFP FRET caspase-2 substrate targeted to mitochondria, we compare FRET efficiencies measured using sensitized emission, acceptor photobleaching and FLI.