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. 2020 Jul 28;10:12568. doi: 10.1038/s41598-020-69412-1

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© The Author(s) 2020

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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Temperature dependence on long-term fluorescent calcium imaging in mammalian cells. (A) HEK cells are known to express temperature-dependent calcium channels, which may trigger calcium in- or efflux through so-called TRP channel families. (B) Image processing workflow shows how we continuously captured fluorescent images using the digital microscope in our incubator setting for single- and multi-cell spatiotemporal calcium fluorometry and metabolic studies. (C1-4) Temperature-dependent changes in intracellular calcium levels in (C1–C2) HEK cells and (C3–C4) primary cortical neurons (E18, rat). (D1–D4) Calcium influx and efflux characteristics show higher temperature sensitivity of HEK cells versus neuronal cells.