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Sci Rep. 2015 Nov 13;5:16487. doi: 10.1038/srep16487.

CalQuo: automated, simultaneous single-cell and population-level quantification of global intracellular Ca2+ responses.

Author information

1
MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, OX3 9DS Oxford, United Kingdom.
2
Department of Chemistry, University of Cambridge, CB2 1EW Cambridge, United Kingdom.
3
Wolfson Imaging Centre Oxford, Weatherall Institute of Molecular Medicine, University of Oxford, OX3 9DS Oxford, United Kingdom.

Abstract

Detecting intracellular calcium signaling with fluorescent calcium indicator dyes is often coupled with microscopy techniques to follow the activation state of non-excitable cells, including lymphocytes. However, the analysis of global intracellular calcium responses both at the single-cell level and in large ensembles simultaneously has yet to be automated. Here, we present a new software package, CalQuo (Calcium Quantification), which allows the automated analysis and simultaneous monitoring of global fluorescent calcium reporter-based signaling responses in up to 1000 single cells per experiment, at temporal resolutions of sub-seconds to seconds. CalQuo quantifies the number and fraction of responding cells, the temporal dependence of calcium signaling and provides global and individual calcium-reporter fluorescence intensity profiles. We demonstrate the utility of the new method by comparing the calcium-based signaling responses of genetically manipulated human lymphocytic cell lines.

PMID:
26563585
PMCID:
PMC4643230
DOI:
10.1038/srep16487
[Indexed for MEDLINE]
Free PMC Article

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