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. 2023 Oct 26;12(21):2526. doi: 10.3390/cells12212526

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Methods to assess calcium handling in cardiomyocytes. Commonly used methods to study calcium signaling in cardiomyocytes include (1) calcium-sensitive dyes and (2) genetically encoded calcium indicators (GECIs). (1) Calcium-sensitive dyes such as Fluo-4 and Fura-2 enter cells and their acetomethoxy (AM) ester moiety is cleaved, allowing the compounds to fluoresce in response to calcium. (2) GECIs are expressed by cells. (2a) GCaMP is composed of circularly permutated enhanced GFP (cpEGFP), calcium-binding domain calmodulin, and calmodulin target sequence M13. Calcium binding to calmodulin increases fluorescence intensity. (2b) RGECO-TnT is composed of the calcium indicator RGECO and troponin T to localize the GECI to the myofilament in order to assess calcium signalling at the site of contraction.