Photo-activatable probes for the analysis of receptor function in living cells.
Photo-activatable (caged) probes are powerful research tools for biological investigation. The superb maneuverability of a light beam allows researchers to activate caged probes with pinpoint accuracy. Recent developments in caging chemistry and two-photon excitation technique further enhance our capability to perform photo-uncaging with even higher spatial and temporal resolution, offering new photonic approaches to study cell signaling dynamics in greater detail. Here we present a sample method that combines the techniques of photo-activation and digital fluorescence microscopy to assay an important class of intracellular receptors for the second messenger D-myo-inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3), or IP(3)). The imaging assay is performed in fully intact living cells using a caged and cell membrane permeable ester derivative of IP(3), cm-IP(3)/PM.
Li
Wen-Hong
WH
Departments of Cell Biology and Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, USA.
D000814:Aniline Compounds / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000737:chemistry / Q000378:metabolism; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000378:metabolism*; D008027:Light; D018613:Microscopy, Confocal; D015335:Molecular Probes / Q000737:chemistry / Q000378:metabolism*; D014966:Xanthenes / Q000378:metabolism
D000814:Aniline Compounds; D053496:Inositol 1,4,5-Trisphosphate Receptors; D015335:Molecular Probes; D014966:Xanthenes; C059715:Fluo-3; D015544:Inositol 1,4,5-Trisphosphate