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Cytosolic Calcium Ions in Melanoma Cancer Cells

Measuring Ca²⁺ with ratiometric fluorescence microscopy

Cytosolic Ca2+ measurement using ratiometric Fura 2 dye Cytosolic_calcium_ions_in_melanoma_cancer_cells_teaser.jpg

In this article, a ratiometric fluorometric method for cytosolic calcium ion (Ca2+) measurement in cultured melanoma cells using Fura 2-AM cell loading and fluorescence microscopy imaging is presented in detail. Cytosolic Ca2+ levels are maintained at low nanomolar concentrations and disruption of Ca2+ homeostasis is associated with cell/tissue damage. Methods have been developed to accurately assess cellular Ca2+ levels, where each has intrinsic advantages and disadvantages. This Fura 2 ratiometric Ca2+ measurement minimizes possible artifacts. The protocol allows evaluation of the immediate effect of chemical addition. Results for this work were obtained using a Leica widefield inverted and fluorescence microscope. For more details, refer to figure 1 and the publication.

Figure 1: The 4 steps of the ratiometric fluorometric method for cytosolic Ca²⁺ measurement in cultured melanoma cells using Fura 2-AM and fluorescence microscopy.

Read the full article:

T Rodrigues, L Silva Ferraz:

Cytosolic Ca2+ measurements by ratiometric fluorescence microscopy in melanoma cells

STAR Protocols (2021) vol. 2, iss. 1, 100282, DOI: 10.1016/j.xpro.2020.100282.

https://www.sciencedirect.com/science/article/pii/S2666166720302690

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