Open AccessImaging Protein‐Protein Interactions by Förster Resonance Energy Transfer (FRET) Microscopy in Live Cells
Author(s) -
Brzostowski Joseph A.,
Meckel Tobias,
Hong Jiang,
Chen Alice,
Jin Tian
Publication year - 2009
Publication title -
current protocols in protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.409
H-Index - 32
eISSN - 1934-3663
pISSN - 1934-3655
DOI - 10.1002/0471140864.ps1905s56
Subject(s) - förster resonance energy transfer , photobleaching , fluorophore , microscopy , acceptor , confocal , confocal microscopy , microscope , live cell imaging , fluorescence , fluorescence lifetime imaging microscopy , fluorescence microscope , chemistry , biophysics , nanotechnology , materials science , optics , physics , cell , biology , biochemistry , condensed matter physics
This unit describes an acceptor‐sensitized emission FRET method using a confocal microscope for image acquisition. In contrast to acceptor photobleaching, which is an end‐point assay that destroys the acceptor fluorophore, the sensitized emission method is amenable for FRET measurements in live cells and can be used to measure changes in FRET efficiency over time. The purpose of this unit is to provide a basic starting point for understanding and performing the sensitized emission method with a simple teaching tool for live‐cell imaging. References that discuss the vagaries of acquiring and analyzing FRET between individually tagged molecules are provided. Curr. Protoc. Protein Sci . 56:19.5.1‐19.5.12. © 2009 by John Wiley & Sons, Inc.