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The understanding of cellular Cd 2+ accumulation and toxicity is hampered by a lack of fluorescent indicators selective for intracellular free Cd 2+ ([Cd 2+ ] i ). In this study, we used depolarized MIN6 mouse pancreatic beta cells as a model for evaluating [Cd 2+ ] i detection with commercially available fluorescent probes, most of which have been traditionally used to visualize [Ca 2+ ] i and [Zn 2+ ] i . We trialed a panel of 12 probes including fura-2, FluoZin-3, Leadmium Green, Rhod-5N, indo-1, Fluo-5N, and others. We found that the [Zn 2+ ] i probe FluoZin-3 and the traditional [Ca 2+ ] i probe fura-2 responded most consistently and robustly to [Cd 2+ ] i accumulation mediated by voltage-gated calcium channels. While selective detection of [Cd 2+ ] i by fura-2 required the omission of Ca 2+ from extracellular buffers, FluoZin-3 responded to [Cd 2+ ] i similarly in the presence or absence of extracellular Ca 2+ . Furthermore, we showed that FluoZin-3 and fura-2 can be used together for simultaneous monitoring of [Ca 2+ ] i and [Cd 2+ ] i in the same cells. None of the other fluorophores tested were effective [Cd 2+ ] i detectors in this model.

Using FluoZin-3 and fura-2 to monitor acute accumulation of free intracellular Cd2+ in a pancreatic beta cell line