Protocol Cytosolic Ca2+ measurements by ratiometric fluorescence microscopy in melanoma cells

Author(s)

Rodrigues, Tiago ; Ferraz, Leticia Silva

• Document type: Journal article
• Source: STAR PROTOCOLS; v. 2, n. 1, p. 12-pg., 2021-03-19.

Abstract

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 ratio -metric fluorometric method for cytosolic Ca2+ measurement in cultured mela-noma 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). (AU)

• FAPESP's process: 18/25747-5 - Alterations of mitochondrial morphology and dynamics in cancer: understanding of tumor biology and prospecting new therapeutic targets
• Grantee: Tiago Rodrigues
• Support Opportunities: Regular Research Grants