Thomas Roger C, Bers Donald M
Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EG, United Kingdom.
Cold Spring Harb Protoc. 2013 Apr 1;2013(4):305-9. doi: 10.1101/pdb.top066290.
It is widely agreed that the best method for measuring the ionized free calcium concentration ([Ca(2+)]) in large volumes of biological solutions is to use Ca(2+)-sensitive macroelectrodes. These are commercially available. To measure [Ca(2+)] in small volumes of solution, minielectrodes with 1-2-mm tips can easily be made and used, and may also be commercially available. Ca(2+)-sensitive microelectrodes (CaSMs, with 0.5-2-μm tips) can also be made and used extracellularly or intracellularly in robust cells, but interest in their use has recently been largely eclipsed. This is because of practical difficulties and the introduction of a large number of fluorescent and other optical calcium probes with calcium sensitivities varying from the nanomolar to the millimolar range, such as Fura-2, Indo-1, Fluo-4, and many others. In this article, we emphasize the utility of Ca(2+)-selective electrodes and show that their use is complementary to use of fluorescent and other optical methods. Each method has advantages and disadvantages. Because numerous reviews and books have been dedicated to the theoretical aspects of ion-selective electrode principles and technology, this article is mainly intended for investigators who have some degree of electrophysiological experience with ion-selective electrodes or microelectrodes.
