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. 2019 Sep 16;9:13307. doi: 10.1038/s41598-019-49791-w

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© The Author(s) 2019

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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Response of a cardiomyocyte (a) to the stimulation by a sequence of pulses. The cardiomyocyte was excited by changes in voltage V_stim between the pipette tips applied as short rectangular pulses ((b), top). The pulse sequence consisted of 5 larger single pulses (pre-pulses, used to precondition the cell) followed by a studied stimulation sequence with varying pulse parameters. In (b) (top), changes in applied voltage are visible with the number of pulses per stimulation sequence marked as Roman numerals on the top of the trace. In response to the pulse, the cardiomyocyte was either stimulated or not. When stimulated, the fluorescence increase was somewhat delayed after the applied stimulation sequence (c), in agreement with the mechanism of Ca²⁺ -induced Ca²⁺ -release. Stimulation was assessed by recording fluorescence F of the Ca²⁺ sensitive dye and is shown normalized to the resting cellular fluorescence F₀. Note that while the preconditioning pulses always elicited a Ca²⁺ transient, the success of cellular activation was variable for the studied pulses. As examples, stimulated (d) and non-stimulated (e) cases are shown with the preceding preconditioning pulse (see subplot markings at the top of the trace in (b) next to the corresponding pulse). In the figure, voltage and fluorescence traces are shown in blue and orange, respectively.