用于人源干细胞衍生心肌细胞的近红外探针的全光学电生理学多模式共轴平台。
Multimodal on-axis platform for all-optical electrophysiology with near-infrared probes in human stem-cell-derived cardiomyocytes.
机构信息
Department of Biomedical Engineering, The George Washington University, Washington, DC, 20052, USA.
Department of Chemistry, University of California, Berkeley, CA, 94720, USA.
出版信息
Prog Biophys Mol Biol. 2020 Aug;154:62-70. doi: 10.1016/j.pbiomolbio.2019.02.004. Epub 2019 Mar 5.
Combined optogenetic stimulation and optical imaging permit scalable, contact-free high-throughput probing of cellular electrophysiology and optimization of stem-cell derived excitable cells, such as neurons and muscle cells. We report a new "on-axis" configuration (combined single optical path for stimulation and for multiparameter imaging) of OptoDyCE, our all-optical platform for studying human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) and other cell types, optically driven by Channelrhodopsin2 (ChR2). This solid-state system integrates optogenetic stimulation with temporally-multiplexed simultaneous recording of membrane voltage (V) and intracellular calcium ([Ca]) dynamics using a single photodetector. We demonstrate the capacity for combining multiple spectrally-compatible actuators and sensors, including newer high-performance near-infrared (NIR) voltage probes BeRST1 and Di-4-ANBDQBS, to record complex spatiotemporal responses of hiPSC-CMs to drugs in a high-throughput manner.
联合光遗传学刺激和光学成像可实现对细胞电生理学的可扩展、无接触式高通量探测，以及优化干细胞衍生的可兴奋细胞，如神经元和肌肉细胞。我们报告了 OptoDyCE 的一种新的“共轴”配置（刺激和多参数成像的单一光学路径的组合），这是我们用于研究人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）和其他细胞类型的全光学平台，由通道视紫红质 2（ChR2）光学驱动。该固态系统将光遗传学刺激与使用单个光电探测器对膜电压（V）和细胞内钙（[Ca]）动力学进行时间多路复用的同时记录相结合。我们展示了结合多个光谱兼容的执行器和传感器的能力，包括新型高性能近红外（NIR）电压探针 BeRST1 和 Di-4-ANBDQBS，以高通量方式记录 hiPSC-CMs 对药物的复杂时空反应。
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