Combining calcium imaging with other optical techniques

Cold Spring Harb Protoc. 2013 Dec 1;2013(12):1125-31. doi: 10.1101/pdb.top066167.


Ca(2+) imaging is a commonly used approach for measuring Ca(2+) signals at high spatial resolution. The method is often combined with electrode recordings to correlate electrical and chemical signals or to investigate Ca(2+) signals following an electrical stimulation. To obtain information on electrical activity at the same spatial resolution, Ca(2+) imaging must be combined with membrane potential imaging. Similarly, stimulation of subcellular compartments requires photostimulation. Thus, combining Ca(2+) imaging with an additional optical technique facilitates the study of a number of physiological questions. The aim of this article is to introduce some basic principles regarding the combination of Ca(2+) imaging with other optical techniques. We discuss the design of the optics, the design of experimental protocols, the optical characteristics of Ca(2+) indicators used in combination with an optical probe, and the affinity of the Ca(2+) indicator in relation to the type of measurement. This information will enable the reader to devise an optimal strategy for combined optical experiments.

Publication types

  • Review

MeSH terms

  • Calcium / analysis*
  • Calcium Signaling
  • Electrophysiological Phenomena*
  • Membrane Potentials
  • Optical Imaging / methods*
  • Staining and Labeling / methods*


  • Calcium