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. 2017 Aug 22;7:9037. doi: 10.1038/s41598-017-09182-5

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Automated multi-neuron tracking of calcium transients during maturation of neuronal cultures. (A) Representative maximum intensity projections of neurons at DIV8–DIV15 with (left) or without (right) ROI overlays (shown in red). Images on right are leveled. (B) Representative calcium traces for 8 individual neurons are shown for each time point. (C) Representative raster plots show Δ fluorescence traces in the 100 most active ROIs within the optical field for frames 1–250. Data are representative of 4 independent experiments and >50,000 total ROIs.