Quantitative imaging of protein-protein interactions by multiphoton fluorescence lifetime imaging microscopy using a streak camera.
Fluorescence lifetime imaging microscopy (FLIM) using multiphoton excitation techniques is now finding an important place in quantitative imaging of protein-protein interactions and intracellular physiology. Recent developments in multiphoton FLIM methods are reviewed and a novel multiphoton FLIM system using a streak camera is described. An example of a typical application of the system is provided in which the fluorescence resonance energy transfer between a donor-acceptor pair of fluorescent proteins within a cellular specimen is measured.
Krishnan
R V
RV
University of Texas Health Science Center, Department of Cellular and Structural Biology, 7703 Floyd Curl Drive, San Antonio, Texas 78229, USA.
Masuda
A
A
Centonze
V E
VE
Herman
B
B
D000818:Animals; D001426:Bacterial Proteins; D053143:Caspase 2; D020169:Caspases / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D006224:Cricetinae; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D049452:Green Fluorescent Proteins; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D008164:Luminescent Proteins; D046911:Macromolecular Substances; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation / Q000379:methods*; D018715:Microscopy, Video / Q000295:instrumentation / Q000379:methods*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins; D020122:tert-Butylhydroperoxide; D053143:Caspase 2; D020169:Caspases