FRET Microscopy for Real-Time Visualization of Second Messengers in Living Cells.
Förster Resonance Energy Transfer (FRET) microscopy is a useful tool in molecular biology and medical research to monitor and quantify real-time dynamics of protein-protein interactions and biochemical processes. Using this well-established technique, many novel signaling mechanisms can be investigated in intact cells or tissues and even in various subcellular compartments. Here, we describe how to perform FRET measurements in living cells expressing FRET-based biosensors and how to evaluate these data. This general protocol can be applied for FRET measurements with various fluorescent biosensors.
Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Martinistr. 52, 20246, Hamburg, Germany.
Axel E
AE
Kraft
Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Martinistr. 52, 20246, Hamburg, Germany. v.nikolaev@uke.de.
Viacheslav O
VO
Nikolaev
D015374:Biosensing Techniques / Q000379:methods; D002460:Cell Line; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D015290:Second Messenger Systems; D015398:Signal Transduction; D013223:Statistics as Topic / Q000379:methods
Biosensor; FRET; Fluorescence; Imaging; Microscopy