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. 2019 Jul 24;9:10716. doi: 10.1038/s41598-019-47009-7

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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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Chemical transfection of the embryonic heart. (A) Diagram of reagents tested for in vivo cardiac transfection. (B) Reagents were microinjected into the pericardial space of windowed HH16 embryos, eggs were then sealed and incubated to desired stages. (C) Representative images of hearts isolated 16 hrs. post transfection with each of the tested reagents. (D) Higher magnification image of an atrial myocyte expressing the palmEGFP 16 hrs. post transfection with Lipofectamine 3000. (E) Volumetric reconstruction of the cell indicated in (D). (F) in ovo image of an embryo 16 hrs. post transfection demonstrating that pericardial injection of Lipofectamine 3000/plasmid DNA reagent preferentially targets the heart (black arrowhead). (G) Comparison of electrical activity in control vs Lipofectamine 3000 transfected hearts. Isochronal maps are drawn at 2 ms/div. Outflow tracts (white dashed lines) were removed prior to imaging. (H) Atrial and ventricular wave traces (dF/F0) from untransfected heart. (I) Atrial and ventricular wave traces (dF/F0) from heart transfected with Lipofectamine 3000. (J) Comparison of Action Potential Duration (APD) between control and transfected hearts (n = 6 per condition). (K) Comparison of total activation time (Ventricular dV/dT max − Atrial dV/dT max) between control and transfected hearts (n = 6 per condition).