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Tytuł:
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Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.
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Autorzy:
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Seibertz F; Institute of Pharmacology and Toxicology, University Medical Center Goettingen, Goettingen, Germany; DZHK (German Center for Cardiovascular Research), Partner Site Goettingen, Germany.
Reynolds M; Cairn Research Ltd, Faversham, United Kingdom.
Voigt N; Institute of Pharmacology and Toxicology, University Medical Center Goettingen, Goettingen, Germany; DZHK (German Center for Cardiovascular Research), Partner Site Goettingen, Germany; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), University of Goettingen, Germany; .
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Źródło:
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Journal of visualized experiments : JoVE [J Vis Exp] 2020 Dec 22 (166). Date of Electronic Publication: 2020 Dec 22.
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Typ publikacji:
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Journal Article; Research Support, Non-U.S. Gov't; Video-Audio Media
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Język:
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English
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Imprint Name(s):
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Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]-
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MeSH Terms:
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Single-Cell Analysis*
Action Potentials/*physiology
Induced Pluripotent Stem Cells/*cytology
Myocytes, Cardiac/*cytology
Cell Differentiation ; Cells, Cultured ; Coloring Agents/chemistry ; Data Analysis ; Electricity ; Humans ; Image Processing, Computer-Assisted
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Substance Nomenclature:
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0 (Coloring Agents)
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Entry Date(s):
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Date Created: 20210111 Date Completed: 20210226 Latest Revision: 20210226
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Update Code:
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20240105
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DOI:
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10.3791/61890
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PMID:
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33427238
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Conventional intracellular microelectrode techniques to quantify cardiomyocyte electrophysiology are extremely complex, labor intensive, and typically carried out in low throughput. Rapid and ongoing expansion of induced pluripotent stem cell (iPSC) technology presents a new standard in cardiovascular research and alternate methods are now necessary to increase throughput of electrophysiological data at a single cell level. VF2.1Cl is a recently derived voltage sensitive dye which provides a rapid single channel, high magnitude response to fluctuations in membrane potential. It possesses kinetics superior to those of other existing voltage indicators and makes available functional data equivalent to that of traditional microelectrode techniques. Here, we demonstrate simplified, non-invasive action potential characterization in externally paced human iPSC derived cardiomyocytes using a modular and highly affordable photometry system.
