4.06 - Fluorescence Resonance Energy Transfer Probes

Abstract

The detection of interactions between molecules is of great interest for the elucidation of cellular functions and mechanisms of disease. Advances in organic and inorganic chemistry and genetics facilitate the production of sophisticated fluorescing substances with defined spectral features, which enable energy transfer provided that they are in close proximity to each other. This mechanism, which was termed ‘fluorescence energy transfer,’ has been enthusiastically used to measure molecular interactions between genes, proteins, etc. in cells. Initially, the methodology was restricted to in vitro studies, but now it extends to the in vivo situation with the emergence of intravital microscopy devices. Researchers are also tempted to transfer the technology to imaging whole living animals noninvasively. This chapter elucidates the principle of fluorescence resonance energy transfer (FRET) and the design of FRET pairs. Intra- and intermolecular FRET applications are also discussed and different methodologies to detect FRET elucidated.