George Washington University, Department of Biomedical Engineering, Washington, DC, United States.
J Biomed Opt. 2023 Jan;28(1):016001. doi: 10.1117/1.JBO.28.1.016001. Epub 2023 Jan 10.
All-optical cardiac electrophysiology enables the visualization and control of key parameters relevant to the detection of cardiac arrhythmias. Mapping such responses in human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) is of great interest for cardiotoxicity and personalized medicine applications.
We introduce and validate a very low-cost compact mapping system for macroscopic all-optical electrophysiology in layers of hiPSC-CMs.
The system uses oblique transillumination, low-cost cameras, light-emitting diodes, and off-the-shelf components (total ) to capture voltage, calcium, and mechanical waves under electrical or optical stimulation.
Our results corroborate the equivalency of electrical and optogenetic stimulation of hiPSC-CMs, and similarity in conduction under pacing. Green-excitable optical sensors are combinable with blue optogenetic actuators (chanelrhodopsin2) only under very low green light ( ). Measurements in warmer culture medium yield larger spread of action potential duration and higher conduction velocities compared to Tyrode's solution at room temperature.
As multiple optical sensors and actuators are combined, our results can help handle the "spectral congestion" and avoid parameter distortion. We illustrate the utility of the system for uncovering the action of cellular uncoupling agents and show extensibility to an epi-illumination mode for future imaging of thicker native or engineered tissues.
全光学心脏电生理学使可视化和控制与检测心律失常相关的关键参数成为可能。在人类诱导多能干细胞衍生的心肌细胞(hiPSC-CMs)中绘制这些反应图对于心脏毒性和个性化医疗应用非常重要。
我们引入并验证了一种非常低成本的紧凑型映射系统,用于 hiPSC-CMs 层中的宏观全光学电生理学。
该系统使用斜向透射、低成本相机、发光二极管和现成组件(总计 )在电或光刺激下捕获电压、钙和机械波。
我们的结果证实了 hiPSC-CMs 的电刺激和光遗传学刺激等效,起搏下的传导相似。只有在绿光( )非常低的情况下,绿色激发光学传感器才能与蓝色光遗传学致动器(channelrhodopsin2)组合使用。与室温下的 Tyrode 溶液相比,在较暖的培养基中测量会导致动作电位持续时间的传播范围更大,传导速度更高。
随着多个光学传感器和致动器的组合,我们的结果可以帮助处理“光谱拥挤”并避免参数失真。我们说明了该系统在揭示细胞去耦剂作用方面的实用性,并展示了对未来更厚的天然或工程组织的 epi-illumination 模式的可扩展性。