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. 2018:1843:69-82.
doi: 10.1007/978-1-4939-8704-7_6.

Fluorescence-Based Measurements of the CRAC Channel Activity in Cell Populations

Pedro C Redondo  1 , Alejandro Berna-Erro  1 , Natalia Dionisio  1 , Juan A Rosado  2
Affiliations

Fluorescence-Based Measurements of the CRAC Channel Activity in Cell Populations

Pedro C Redondo et al. Methods Mol Biol. 2018.

Abstract

Cytosolic Ca2+ plays an important role in cellular biology, and since its identification as a second messenger, a number of techniques and methods to analyze the changes in cytosolic Ca2+ concentration ([Ca2+]c) induced by physiological agonists have been developed. Changes in [Ca2+]c might be determined in single cells or in cell populations. Measurement in single cells allows to determine changes in [Ca2+]c at a subcellular level but often results in heterogeneous responses among cells. Determination of intracellular Ca2+ mobilization at the cell population level reduces this heterogeneity and allows [Ca2+]c measurements in small cells that load little amounts of indicator. Here, we describe the measurement of agonist-evoked changes in [Ca2+]c associated with Ca2+ influx in cell populations.

Keywords: Calcium homeostasis; Cytosolic calcium; ELISA; Flow cytometry; Fluorescence spectrophotometer; Fluorescent probes; Ion channels.

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