Optopatch: high-throughput all-optical electrophysiology

项目摘要 尽管许多神经疾病的流行和严重程度，新的发展 50年来，药品类别一直停滞不前。这在很大程度上是由于缺乏好的模式。 快速研究相关电和突触表型的系统和工具。我们的目标是 克服这些挑战。诱导多能干细胞(IPSC)研究进展 技术揭示了结合临床病史研究人类神经元的第一个前景 使用快速体外技术。这些细胞复杂电生理行为可以是 使用Optopatch平台和Q-State最近开发的显微镜系统进行记录。 使用这些工具，可以同时刺激(蓝光)和记录(红光) 来自大约100个神经元的电活动，时间分辨率为1毫秒， 单细胞空间分辨率，高信噪比。该系统可以用来测量 单细胞兴奋性和放电模式或通过刺激一个亚群来探测突触传递 具有空间图案的蓝光的神经元。展望未来，我们建议增加 在不牺牲性能的情况下放大吞吐量并严格测试平台的 性能。首先，显微镜将升级为先进的环境控制系统，96- 良好的板材兼容性，以及用于复合添加的流体处理机器人。接下来是数据存储 分析将安全地转移到云中，以处理2.5 TB/天的数据速率。一次 显微镜功能齐全，灵敏度高，重现性好(从一块到另一块，从一块到另一块 一批又一批)将使用对照化合物库进行测试。最后，作为第一个应用程序， 我们将寻找一种强健的、可筛选的Drave型综合征的表型。神经元将会是 从10名健康的患者和10名德雷特患者身上准备的，以寻找超越 遗传背景的变异。一种能改善人类疾病表型的药物 大多数细胞系是临床上有希望广泛有效的候选细胞。一口井 经过验证的高通量电生理平台，具有确认的表型读数 人类IPSC病神经元有可能改变药物筛选格局 神经紊乱。我们希望开辟一条新的途径来治疗这些可怕的疾病 疾病。

Project Summary In spite of the prevalence and severity of many neurological disorders, the development of new classes of drugs has been sluggish for 50 years. This is due largely to the lack of good model systems and tools to rapidly study relevant electrical and synaptic phenotypes. We aim to overcome these challenges. Recent advances in induced pluripotent stem cell (iPSC) technology reveal the first prospects for studying human neurons paired with clinical histories using fast in vitro technologies. The complex electrophysiological behavior of these cell can be recorded with the Optopatch platform and microscope systems recently developed at Q-State. With these tools, it is possible to simultaneously stimulate (blue light) and record (red light) electrical activity from around a hundred neurons with one millisecond temporal resolution, single cell spatial resolution, and high signal to noise. This system can be used to measure single cell excitability and firing patters or to probe synaptic transmission by stimulating a subset of neurons with spatially patterned blue light. Moving forward, we propose to increase microscope throughput without sacrificing capabilities and rigorously test the platform’s performance. First, the microscope will be upgraded with advanced environmental controls, 96- well plate compatibility, and a fluid-handling robot for compound addition. Next, data storage and analysis will be securely moved to the cloud to handle 2.5 Terabyte/day data rates. Once the microscope is fully functional, sensitivity and reproducibility (well to well, plate to plate, and batch to batch) will be tested using a library of control compounds. Finally, as a first application, we will search for a robust, screenable phenotype for Dravet syndrome. Neurons will be prepared from ten healthy and ten Dravet patients to look for differences in firing that transcend variation in the genetic background. A drug that ameliorates the disease phenotype in the majority of cell lines is a promising candidate to be broadly effective in the clinic. A well validated, high-throughput electrophysiology platform with confirmed phenotypic readouts in human iPSC disease neurons has the potential to change the drug screening landscape for neurological disorders. We hope to open a new path to finding treatments for these horrible diseases.