Quantitative intensity-based FRET approaches--a comparative snapshot.

摘要: Forster resonance energy transfer (FRET) has become an important tool for analyzing different aspects of interactions among biological macromolecules in their native environments. FRET analysis also been successfully applied to study the spatiotemporal regulation various cellular processes using genetically encoded FRET-based biosensors. A variety procedures have described measuring efficiency or relative abundance donor-acceptor complexes, based on donor fluorescence lifetime spectrally resolved intensity. The latter methods are preferable if one wants not only quantify apparent efficiencies but calculate stoichiometry and observe fast dynamic changes acceptor molecules live cells. This review focuses a comparison available intensity-based approaches used measure FRET. We discuss strengths weaknesses terms quantification, provide several examples applications.