Imaging single-channel calcium microdomains by total internal reflection microscopy.
The microdomains of Ca2+ in the cytosol around the mouth of open Ca2+ channels are the basic 'building blocks' from which cellular Ca2+ signals are constructed. Moreover, the kinetics of local [Ca2+] closely reflect channel gating, so their measurement holds promise as an alternative to electrophysiological patch-clamp recording as a means to study single channel behavior. We have thus explored the development of optical techniques capable of imaging single-channel Ca2+ signals with good spatial and temporal resolution, and describe results obtained using total internal reflection fluorescence microscopy to monitor Ca2+ influx through single N-type channels expressed in Xenopus oocytes.
Demuro
Angelo
A
Department of Neurobiology and Behavior, University of California, Irvine, CA 92697, USA.
Parker
Ian
I
D000818:Animals; D015220:Calcium Channels / Q000378:metabolism*; D004553:Electric Conductivity; D004594:Electrophysiology; D015640:Ion Channel Gating; D007700:Kinetics; D021962:Membrane Microdomains; D008564:Membrane Potentials; D008856:Microscopy, Fluorescence / Q000379:methods*; D009865:Oocytes / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D014982:Xenopus laevis
D015220:Calcium Channels