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

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


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aniline Compounds / chemistry
  • Calcium / chemistry
  • Calcium / metabolism*
  • Calcium Signaling
  • Cell Line, Tumor
  • Computational Biology / methods*
  • Fluorescent Dyes / chemistry
  • HEK293 Cells
  • Humans
  • Intracellular Space / metabolism*
  • Jurkat Cells
  • Reproducibility of Results
  • Single-Cell Analysis / methods*
  • Software*
  • T-Lymphocytes / metabolism
  • Time-Lapse Imaging / methods
  • Xanthenes / chemistry


  • Aniline Compounds
  • Fluo 4
  • Fluorescent Dyes
  • Xanthenes
  • Calcium