Converting fluorescence data into Ca²⁺ concentration

Bootman, Martin D.; Rietdorf, Katja; Collins, Tony; Walker, Simon and Sanderson, Michael (2013). Converting fluorescence data into Ca²⁺ concentration. Cold Spring Harbor Protocols, 2013(2) pp. 126–129.

DOI: https://doi.org/10.1101/pdb.prot072827

Abstract

In many situations, fluorescent Ca²⁺ reporters are used to simply indicate that a change of Ca²⁺ concentration has occurred. Monitoring the emission from a Ca²⁺-sensitive indicator can be sufficient to tell whether a signal has arisen, and what its kinetic/spatial parameters were. The emission from an indicator does not have a linear relationship to the Ca²⁺ concentration within a cell; rather, the relationship between fluorescence emission and Ca²⁺ concentration is described by a logistic function. Simply recording fluorescence emission, therefore, provides a relative indication of the magnitude of a Ca²⁺ signal that should not be used for generating mean amplitude data. However, with a little consideration and effort, the fluorescence output can be calibrated to yield actual Ca²⁺ concentration.

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