Imaging protein molecules using FRET and FLIM microscopy.
Förster (or fluorescence) resonance energy transfer (FRET) and fluorescence lifetime imaging (FLIM) have moved center stage and are increasingly forming part of multifaceted imaging approaches. They are complementary methodologies that can be applied to advanced quantitative analyses. The widening application of FRET and FLIM has been driven by the availability of suitable fluorophores, increasingly sophisticated microscopy systems, methodologies to correct spectral bleed-through, and the ease with which FRET can be combined with other techniques. FRET and FLIM have recently found use in several applications: in the analysis of protein-protein interactions with high spatial and temporal specificity (e.g. clustering), in the study of conformational changes, in the analysis of binding sequences, and in applications such as high-throughput screening.
Wallrabe
Horst
H
Keck Center for Cellular Imaging, Department of Biology, University of Virginia, Gilmer Hall, Charlottesville, Virginia 22904, USA.
Periasamy
Ammasi
A
D031541:Fluorescence Resonance Energy Transfer / Q000379:methods* / Q000639:trends; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*
D011506:Proteins