Photobleaching-corrected FRET efficiency imaging of live cells.

ABSTRACT: Fluorescent resonance energy transfer (FRET) imaging techniques can be used to visualize protein-protein interactions in real-time with subcellular resolution. Imaging of sensitized fluorescence of the acceptor, elicited during excitation of the donor, is becoming the most popular method for live FRET (3-cube imaging) because it is fast, nondestructive, and applicable to existing widefield or confocal microscopes. Most sensitized emission-based FRET indices respond nonlinearly to changes in the degree of molecular interaction and depend on the optical parameters of the imaging system. This makes it difficult to evaluate and compare FRET imaging data between laboratories. Furthermore, photobleaching poses a problem for FRET imaging in timelapse experiments and three-dimensional reconstructi

SUBMITTER: Zal T 

PROVIDER: S-EPMC1304294 | biostudies-literature | 2004 Jun

REPOSITORIES: biostudies-literature