人类多能干细胞衍生心肌细胞中电压和钙传感器双光学映射的实验方案。
Protocol for dual-optical mapping of voltage and calcium sensors in human pluripotent stem cell-derived cardiomyocytes.
作者信息
Alibhai Faisal J, Masoumi Arya, Kim Ho-June, Kim Nathan, Kim Wesley, Gomez-Garcia Juliana, Lee Peter, Laflamme Michael A
机构信息
McEwen Stem Cell Institute, University Health Network, Toronto, ON, Canada.
Essel Research and Development Inc., Toronto, ON, Canada; Faculty of Science, McMaster University, Hamilton, ON, Canada.
出版信息
STAR Protoc. 2025 Aug 23;6(3):104044. doi: 10.1016/j.xpro.2025.104044.
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.
同时获取荧光电压和细胞内钙信号需要光谱不同的指示剂和高速成像装置。在这里，我们提出了一种用于在人多能干细胞衍生的心肌细胞（hPSC-CM）单层中对这些关键生理参数进行光学映射的方案。我们描述了使用涉及荧光染料和基因编码传感器的光谱兼容电压和钙传感器的步骤。我们还详细介绍了构建一个在图像采集期间能够与高速相机同步控制发光二极管（LED）的驱动器的程序。
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