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Measurement of [Ca2+]i in Cell Suspensions Using Indo-1

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Intracellular calcium, in particular the cytosolic free ionized calcium concentration [Ca2+ ]i , is tightly regulated under physiological conditions. Stimulation of receptors, belonging to almost all the classes so far described, will result in changes in [Ca2+ ]i . These changes might be directly induced by either Ca2+ -influx or Ca2+ -mobilization from intracellular stores, or indirectly by a number of other mechanisms (1 4 ). The development of fluorescent indicators of free [Ca2+ ] that could be loaded into intact cells has contributed enormously to the understanding of cellular Ca2+ homeostasis, especially dyes that respond to Ca2+ with shifts of excitation or emission spectra (5 7 ). By measuring at two selected wavelengths (either dual emission or dual excitation), it is possible to calculate the proportion of dye in the Ca2+ -bound and Ca2+ -free forms.
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