Barcoding T cell calcium response diversity with methods for automated and accurate analysis of cell signals (MAAACS)

PLoS Comput Biol. 2013;9(9):e1003245. doi: 10.1371/journal.pcbi.1003245. Epub 2013 Sep 26.

Abstract

We introduce a series of experimental procedures enabling sensitive calcium monitoring in T cell populations by confocal video-microscopy. Tracking and post-acquisition analysis was performed using Methods for Automated and Accurate Analysis of Cell Signals (MAAACS), a fully customized program that associates a high throughput tracking algorithm, an intuitive reconnection routine and a statistical platform to provide, at a glance, the calcium barcode of a population of individual T-cells. Combined with a sensitive calcium probe, this method allowed us to unravel the heterogeneity in shape and intensity of the calcium response in T cell populations and especially in naive T cells, which display intracellular calcium oscillations upon stimulation by antigen presenting cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Calcium / metabolism*
  • Chlorocebus aethiops
  • Humans
  • Molecular Probes
  • Signal Transduction*
  • Software*
  • T-Lymphocytes / metabolism*

Substances

  • Molecular Probes
  • Calcium

Grants and funding

ASa's PhD thesis was supported by fellowships from the French “Ministère de la Recherche” and from the ARC (Association pour la Recherche contre le Cancer). MCP was funded by FRM (Fondation pour la recherche médicale, France). CB was funded by the Agence Nationale de la Recherche (ANR Blanc-Nanodigicode). This work was also funded by a specific grant from the Agence Nationale de la Recherche (ANR-10-BLAN-1509 and ANR-10-INBS-04-01 France Bio Imaging). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.