Measuring Cytoplasmic Calcium in Single Living Cells Using Fluorescent Probes

  • Ron Jacob
  • Christopher D. Benham
Part of the Topics in Molecular and Structural Biology book series (TMSB)


Calcium is an important intracellular second messenger in that it often links events occurring at the cell surface, such as a membrane potential depolarization or the binding of a hormone to a receptor, with events occurring within the cell, such as activation of the contractile apparatus or secretion. The importance of calcium was recognized long before it was possible to measure it at the very low concentrations that occur intracellularly, but an improved understanding of its role developed when its direct measurement became feasible. However, the earlier techniques for measuring cytoplasmic free calcium ion concentration ([Ca2+]j) not only were technically difficult, but also were confined to use with large cells (e.g. squid giant axon or large muscle cells) because of the need to introduce some form of micropipette into the cell, either to microinject the indicator or as part of a Ca2+-sensitive microelectrode (but see Campbell et al., 1985, for a review of alternative strategies for introducing photoproteins into cells). The introduction of the fluorescent indicators has overcome these technical difficulties, resulting in their widespread use. A very good comprehensive review of optical indicators of [Ca2+] is Blinks et al (1982), although it mentions only the first of the fluorescent indicators (quin2).


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© The Macmillan Press Ltd 1991

Authors and Affiliations

  • Ron Jacob
  • Christopher D. Benham

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