OptoDyCE as an automated system for high-throughput all-optical dynamic cardiac electrophysiology.
OptoDyCE as an automated system for high-throughput all-optical dynamic cardiac electrophysiology.
复制标题
光能作为高通量全光动态心脏电生理学的自动化系统。
DOI：
10.1038/ncomms11542
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发表时间：
2016-05-10
影响因子：
16.6
通讯作者：
Entcheva E
中科院分区：
文献类型：
--
作者：
Klimas A;Ambrosi CM;Yu J;Williams JC;Bien H;Entcheva E
临床前心脏毒性测试的改进，新的离子通道靶向药物的发现，干细胞来源的心肌细胞和其他生物制剂的表型和使用都需要高通量（HT），细胞水平的电生理审讯工具。用于驱动和传感的光学技术提供了即时并行性，使细胞和小组织结构的非接触动态高温测试成为可能，这是其他方法无法负担得起的。在这里，我们通过计算和实验展示了全光电生理学在药物测试中的局限性，然后实现并验证了OptoDyCE，一个全光心脏电生理学的全自动系统。我们通过病毒式地在大鼠和人类心肌细胞中引入光遗传驱动程序或通过专用光敏体细胞“火花”细胞的模块化使用来验证光学驱动。我们表明，这种自动化的全光学方法提供了细胞询问的HT手段，即允许每小时动态测试bb60 600多细胞样品或化合物，并产生关于药物随时间、空间和剂量的作用的高含量信息。通过使用光遗传学工具，可以提高临床前药物测试和细胞功能表征的效率。在这里，Klimas等人展示并验证了OptoDyCE，一种全光高通量心脏电生理全自动系统。
The improvement of preclinical cardiotoxicity testing, discovery of new ion-channel-targeted drugs, and phenotyping and use of stem cell-derived cardiomyocytes and other biologics all necessitate high-throughput (HT), cellular-level electrophysiological interrogation tools. Optical techniques for actuation and sensing provide instant parallelism, enabling contactless dynamic HT testing of cells and small-tissue constructs, not affordable by other means. Here we show, computationally and experimentally, the limits of all-optical electrophysiology when applied to drug testing, then implement and validate OptoDyCE, a fully automated system for all-optical cardiac electrophysiology. We validate optical actuation by virally introducing optogenetic drivers in rat and human cardiomyocytes or through the modular use of dedicated light-sensitive somatic ‘spark' cells. We show that this automated all-optical approach provides HT means of cellular interrogation, that is, allows for dynamic testing of >600 multicellular samples or compounds per hour, and yields high-content information about the action of a drug over time, space and doses. The efficiency of preclinical drug testing and characterization of cellular function can be improved through the use of optogenetic tools. Here Klimas et al. present and validate OptoDyCE, a fully automated system for all-optical high-throughput cardiac electrophysiology.