Localizing protein-protein interactions in living cells using fluorescence lifetime imaging microscopy.
In the past decade, advances in fluorescence lifetime imaging have extensively applied in the life sciences, from fundamental biological investigations to advanced clinical diagnosis. Fluorescence lifetime imaging microscopy (FLIM) is now routinely used in the biological sciences to monitor dynamic signaling events inside living cells, e.g., Protein-Protein interactions. In this chapter, we describe the calibration of both time-correlated single-photon counting (TCSPC) and frequency domain (FD) FLIM systems and the acquisition and analysis of FLIM-FRET data for investigating Protein-Protein interactions in living cells.
W.M. Keck Center for Cellular Imaging, Biology, University of Virginia, B005 Physical and Life Sciences Building, White Head Road, Charlottesville, VA, 22904, USA.
Yuansheng
Y
Sun
Ammasi
A
Periasamy
D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D008962:Models, Theoretical; D061848:Optical Imaging / Q000379:methods* / Q000592:standards; D025941:Protein Interaction Mapping / Q000379:methods*