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Methods to Measure Intracellular Ca2+ Concentration Using Ca2+-Sensitive Dyes

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Calcium Signalling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1925))

Abstract

Ca2+ ion is universally considered the most versatile second messenger responsible for decoding and regulating the majority of the signaling pathways within the cell. The study of intracellular Ca2+ concentration ([Ca2+]i) dynamics is consequently of primary importance for the interpretation of cellular biology. This chapter will present a relatively simple, largely diffused, and nevertheless robust method to measure variations of [Ca2+]i by the use of the Ca2+-sensitive chemical dye Fura-2. A general protocol for the assessment of [Ca2+]i in adherent cells, applicable to a variety of cell systems, will be first presented. Then, the implementation of Fura-2 to detect [Ca2+]i in two specific cell types, namely, human adrenocortical cells and primary skin fibroblasts, will be discussed in more particulars. Finally, the procedure to monitor Ca2+ influx through the plasma membrane using Fura-2 will be described.

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Authors and Affiliations

  1. Department of Medicine, University of Padua, Padua, Italy

    Sofia Zanin

  2. Department of Biomedical Sciences, University of Padua, Padua, Italy

    Elisa Lidron, Rosario Rizzuto & Giorgia Pallafacchina

  3. Neuroscience Institute, Italian National Research Council (CNR), Padua, Italy

    Giorgia Pallafacchina

Authors
  1. Sofia Zanin
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  2. Elisa Lidron
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  3. Rosario Rizzuto
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  4. Giorgia Pallafacchina
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Corresponding author

Correspondence to Giorgia Pallafacchina .

Editor information

Editors and Affiliations

  1. Department of Biomedical Sciences, University of Padua, Padua, Italy

    Anna Raffaello

  2. Department of Biomedical Sciences, University of Padua, Padua, Italy

    Denis Vecellio Reane

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Zanin, S., Lidron, E., Rizzuto, R., Pallafacchina, G. (2019). Methods to Measure Intracellular Ca2+ Concentration Using Ca2+-Sensitive Dyes. In: Raffaello, A., Vecellio Reane, D. (eds) Calcium Signalling. Methods in Molecular Biology, vol 1925. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9018-4_4

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  • DOI: https://doi.org/10.1007/978-1-4939-9018-4_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9017-7

  • Online ISBN: 978-1-4939-9018-4

  • eBook Packages: Springer Protocols

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