Cytosolic Ca²⁺ measurements by ratiometric fluorescence microscopy in melanoma cells

Cytosolic Ca²⁺ levels are maintained at low nanomolar concentrations, and disruption of Ca²⁺ homeostasis is associated with cell/tissue damage. Thus, methods have been developed to accurately assess cellular Ca²⁺ levels, each with intrinsic advantages and disadvantages. Here, we present in detail a ratiometric fluorometric method for cytosolic Ca²⁺ measurement in cultured melanoma cells using Fura 2-AM cell loading and fluorescence microscopy imaging. For complete details on the use and execution of this protocol, please refer to Esteves et al. (2020).