Versatile and automated workflow for the analysis of oligodendroglial calcium signals

Although intracellular Ca 2+ signals of oligodendroglia, the myelin-forming cells of the central nervous system, regulate vital cellular processes including myelination, few studies on oligodendroglia Ca 2+ signal dynamics have been carried out and existing software solutions are not adapted to the analysis of the complex Ca 2+ signal characteristics of these cells. Here, we provide a comprehensive solution to analyze oligodendroglia Ca 2+ imaging data at the population and single-cell levels. We describe a new analytical pipeline containing two free, open source and cross-platform software programs, Occam and post-prOccam, that enable the fully automated analysis of one- and two-photon Ca 2+ imaging datasets from oligodendroglia obtained by either ex vivo or in vivo Ca 2+ imaging techniques. Easily configurable, our software solution is optimized to obtain unbiased results from large datasets acquired with different imaging techniques. Compared to other recent software, our solution proved to be fast, low memory demanding and faithful in the analysis of oligodendroglial Ca 2+ signals in all tested imaging conditions. Our versatile and accessible Ca 2+ imaging data analysis tool will facilitate the elucidation of Ca 2+ -mediated mechanisms in oligodendroglia. Its configurability should also ensure its suitability with new use cases such as other glial cell types or even cells outside the CNS.