Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytes
Summary: Simultaneous acquisition of fluorescent voltage and intracellular calcium signals requires spectrally distinct indicators and a high-speed imaging setup. Here, we present a protocol for the optical mapping of these key physiological parameters in human pluripotent stem cell-derived cardiomy...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725004502 |
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| author | Faisal J. Alibhai Arya Masoumi Ho-June Kim Nathan Kim Wesley Kim Juliana Gomez-Garcia Peter Lee Michael A. Laflamme |
| author_facet | Faisal J. Alibhai Arya Masoumi Ho-June Kim Nathan Kim Wesley Kim Juliana Gomez-Garcia Peter Lee Michael A. Laflamme |
| author_sort | Faisal J. Alibhai |
| collection | DOAJ |
| description | Summary: Simultaneous acquisition of fluorescent voltage and intracellular calcium signals requires spectrally distinct indicators and a high-speed imaging setup. Here, we present a protocol for the optical mapping of these key physiological parameters in human pluripotent stem cell-derived cardiomyocyte (hPSC-CM) monolayers. We describe steps for using spectrally compatible voltage and calcium sensors involving fluorescent dyes and genetically encoded sensors. We also detail procedures for building a driver capable of controlling light-emitting diodes (LEDs) in synchrony with a high-speed camera during image acquisition. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-53e7fcaf4d9b408d993c30b108cf3cc2 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-53e7fcaf4d9b408d993c30b108cf3cc22025-08-24T05:14:34ZengElsevierSTAR Protocols2666-16672025-09-016310404410.1016/j.xpro.2025.104044Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytesFaisal J. Alibhai0Arya Masoumi1Ho-June Kim2Nathan Kim3Wesley Kim4Juliana Gomez-Garcia5Peter Lee6Michael A. Laflamme7McEwen Stem Cell Institute, University Health Network, Toronto, ON, Canada; Corresponding authorEssel Research and Development Inc., Toronto, ON, Canada; Faculty of Science, McMaster University, Hamilton, ON, CanadaEssel Research and Development Inc., Toronto, ON, Canada; Department of Electrical and Computer Engineering, University of Toronto, Toronto, ON, CanadaEssel Research and Development Inc., Toronto, ON, Canada; Department of Computer Science, Stanford University, Stanford, CA, USAEssel Research and Development Inc., Toronto, ON, Canada; The Department of Mechanical and Mechatronics Engineering, University of Waterloo, Waterloo, ON, CanadaMcEwen Stem Cell Institute, University Health Network, Toronto, ON, CanadaMcEwen Stem Cell Institute, University Health Network, Toronto, ON, Canada; Essel Research and Development Inc., Toronto, ON, Canada; Corresponding authorMcEwen Stem Cell Institute, University Health Network, Toronto, ON, Canada; Peter Munk Cardiac Centre, University Health Network, Toronto, ON, Canada; Department of Laboratory Medicine & Pathobiology, University of Toronto, Toronto, ON, Canada; Corresponding authorSummary: Simultaneous acquisition of fluorescent voltage and intracellular calcium signals requires spectrally distinct indicators and a high-speed imaging setup. Here, we present a protocol for the optical mapping of these key physiological parameters in human pluripotent stem cell-derived cardiomyocyte (hPSC-CM) monolayers. We describe steps for using spectrally compatible voltage and calcium sensors involving fluorescent dyes and genetically encoded sensors. We also detail procedures for building a driver capable of controlling light-emitting diodes (LEDs) in synchrony with a high-speed camera during image acquisition. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725004502Health SciencesMolecular/Chemical ProbesStem Cells |
| spellingShingle | Faisal J. Alibhai Arya Masoumi Ho-June Kim Nathan Kim Wesley Kim Juliana Gomez-Garcia Peter Lee Michael A. Laflamme Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytes STAR Protocols Health Sciences Molecular/Chemical Probes Stem Cells |
| title | Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytes |
| title_full | Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytes |
| title_fullStr | Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytes |
| title_full_unstemmed | Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytes |
| title_short | Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytes |
| title_sort | protocol for dual optical mapping of voltage and calcium sensors in human pluripotent stem cell derived cardiomyocytes |
| topic | Health Sciences Molecular/Chemical Probes Stem Cells |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725004502 |
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