Department of Biomedical Engineering, The George Washington University, Washington, DC, 20052, USA.
Department of Chemistry, University of California, Berkeley, CA, 94720, USA.
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。
