Developing nanoscale electrophysiology sensors for robust intracellular recording

项目摘要/摘要： 生电细胞的动作电位，如神经元和心肌细胞，对它们的 生理功能。神经元使用动作电位来远距离传递信号，而心肌细胞 在每一次心跳中，使用动作电位来同步数百万个细胞的收缩。要理解 这些重要的生理功能，其中最重要的策略之一就是准确记录电信号 来自细胞的电势。然而，目前的两大类电生理方法，细胞内和 细胞外记录在其应用中受到严重限制。细胞内记录，如膜片钳 遭受极低的吞吐量和有毒的细胞内透析。细胞外记录，如平面记录 电极阵列信号差，电池与电极之间缺乏一对一的耦合。在过去的十年里， 很多工作都集中在开发新一代电生理工具上，以实现高 吞吐量细胞内记录。特别是，开发了基于纳米技术的电极传感器 独立地在几个组中显示出在实现高度敏感和高吞吐量方面的巨大前景 细胞内记录。然而，将这些新兴技术开发成强大的电生理工具 将需要进行大量的表征、验证和优化研究。这项提案旨在发展 将纳米电极技术转化为生物医学研究的强大电生理工具。完成后， 这项新技术将使用户能够(A)进行敏感的细胞内动作电位记录 同时记录数十至数百个单个细胞；(B)实现长期、微创的记录 细胞数天至数周；以及(C)在最佳环境下提供稳定的HSC-CMS培养和记录 条件。

Project Summary / Abstract: Action potentials of electrogenic cells, such as neurons and cardiomyocytes, are crucial for their physiological functions. Neurons use action potential to transmit signals over long distances, and cardiomyocytes use action potentials to synchronize the contraction of millions of cells during each heartbeat. To understand these important physiological functions, one of the most important tactics is to accurately record the electrical potentials from cells. However, the current two major classes of electrophysiological methods, intracellular and extracellular recording, suffer severe limitations in their applications. Intracellular recording such as patch clamp suffers from extremely low throughput and toxic intracellular dialysis. Extracellular recording such as planar electrode array suffers from poor signal and lack of one-to-one cell-to-electrode coupling. In the last decade, much effort has been focused on developing new generation of electrophysiology tools to achieve high throughput intracellular recording. In particular, nanotechnology-based electrode sensors developed independently in several groups has shown great promise in achieving highly sensitive and high throughput intracellular recording. However, developing these nascent technologies into robust electrophysiological tools would require extensive studies for characterization, validation, and optimization. This proposal aims to develop the nanoelectrode technology into robust electrophysiological tools for biomedical research. When accomplished, this new technology will enable users to (a) perform sensitive, intracellular recording of action potentials from tens to hundreds of individual cells simultaneously; (b) achieve long-term, minimally-invasive recording of the cells for days to weeks; and (c) afford stable culture and recording of the hSC-CMs under optimal environmental conditions.