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Fuente Pérez, Sergio De La 2025-09-16T12:27:32Z 2025-09-16T12:27:32Z 2019 Fernandez-Sanz C, De la Fuente S, Sheu SS. Mitochondrial Ca2+ concentrations in live cells: quantification methods and discrepancies. FEBS Lett. 2019 Jul;593(13):1528-1541. doi: 10.1002/1873-3468.13427. Epub 2019 May 18. PMID: 31058316; PMCID: PMC7573507. https://uvadoc.uva.es/handle/10324/77797 10.1002/1873-3468.13427. Intracellular Ca2+ signaling controls numerous cellular functions. Mitochondria respond to cytosolic Ca2+ changes by adapting mitochondrial functions and, in some cell types, shaping the spatiotemporal properties of the cytosolic Ca2+ signal. Numerous methods have been developed to specifically and quantitatively measure the mitochondrial-free Ca2+ concentrations ([Ca2+ ]m ), but there are still significant discrepancies in the calculated absolute values of [Ca2+ ]m in stimulated live cells. These discrepancies may be due to the distinct properties of the methods used to measure [Ca2+ ]m , the calcium-free/bound ratio, and the cell-type and stimulus-dependent Ca2+ dynamics. Critical processes happening in the mitochondria, such as ATP generation, ROS homeostasis, and mitochondrial permeability transition opening, depend directly on the [Ca2+ ]m values. Thus, precise determination of absolute [Ca2+ ]m values is imperative for understanding Ca2+ signaling. This review summarizes the reported calibrated [Ca2+ ]m values in many cell types and discusses the discrepancies among these values. Areas for future research are also proposed. eng info:eu-repo/semantics/openAccess Mitochondrial Ca2+ concentrations in live cells: quantification methods and discrepancies info:eu-repo/semantics/article