Quantification of cytosolic 'free' calcium in isolated coral cells with confocal microscopy

Despite its prominent role as an intracellular messenger in all organisms, cytosolic free calcium ([Ca2+]i) has never been quantified in corals or cnidarians in general. Ratiometric calcium dyes and cell imaging have been key methods in successful research on [Ca2+]i in model systems, and could be applied to corals. Here, we developed a procedure to quantify [Ca2+]i in isolated cells from the model coral species Stylophora pistillata using Indo-1 and confocal microscopy. We quantified [Ca2+]i in coral cells with and without intracellular dinoflagellate symbionts, and verified our procedure on cultured mammalian cells. We then used our procedure to measure changes in [Ca2+]i in coral cells exposed to a classic inhibitor of [Ca2+]i regulation, thapsigargin, and also used it to record elevations in [Ca2+]i in coral cells undergoing apoptosis. Our procedure paves the way for future studies into intracellular calcium in corals and other cnidarians.