Measurement of Intracellular Calcium Concentration Using Confocal Microscopy
Many cellular functions are tightly regulated by intracellular calcium concentrations ([Ca2+]i), and, therefore, the measurement of [Ca2+]i is of critical importance. To determine Ca2+-related cellular dynamics accurately, it is necessary to measure three-dimensionally resolved [Ca2+]i with sufficient temporal resolution to follow fast cellular responses that generate signal pulses and wave propagation. Several techniques have been developed to assay [Ca2+]i, but a revolution in the study of intracellular [Ca2+]i occurred when fluorescent dyes for Ca2+ were developed (1). Since then, fluorescent dyes and fluorescent microscopy have been used to observe resting and nonresting [Ca2+] in intact cells as well as in subcellular fractions.
KeywordsCage MgCl2 Stratification NADH HEPES
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