Imaging single-channel calcium microdomains by total internal reflection microscopy

Biol Res. 2004;37(4):675-9. doi: 10.4067/s0716-97602004000400025.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Calcium Channels / metabolism*
  • Electric Conductivity
  • Electrophysiology
  • Ion Channel Gating
  • Kinetics
  • Membrane Microdomains*
  • Membrane Potentials
  • Microscopy, Fluorescence / methods*
  • Oocytes / metabolism*
  • Patch-Clamp Techniques
  • Xenopus laevis

Substances

  • Calcium Channels