Label-free Optical Recording of Neuroelectric Activities

神经电活动的无标记光学记录

• 批准号: 10190148
• 负责人: Bianxiao Cui
• 金额: $ 43.41万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10190148
• 关键词: Action Potentials; Area; Axon; Back; Biomedical Engineering; Brain; Calcium; Cardiac Myocytes; Cell physiology; Cell-Free System; Cells; Complex; Computer software; Data; Dendrites; Detection; Dopamine; Dyes; Electrophysiology (science); Film; Fluorescence; Frequencies; Funding Opportunities; Goals; Hippocampus (Brain); Image; Imaging Techniques; Label; Lasers; Location; Measurement; Measures; Methods; Microscope; Midbrain structure; Molecular Probes; Neurons; Neurosciences; Optics; Pacemakers; Photobleaching; Phototoxicity; Process; Property; Proteins; Reaction Time; Research; Research Personnel; Research Project Grants; Resolution; Sampling; Scanning; Shapes; Signal Transduction; Site; Slice; Spottings; System; Techniques; Technology; Thinness; Time; Work; absorption; base; brain tissue; design; dopaminergic neuron; electrical potential; extracellular; flexibility; fluorescence imaging; improved; interest; multi-electrode arrays; neural circuit; novel; optical imaging; patch clamp; response; sensor; stem cells; tool; voltage; voltage sensitive dye

Project Summary Understanding how a network of interconnected neurons receives, stores and processes information requires parallel and high quality recording of neuroelectric signals. Intracellular recording techniques such as patch clamp are invasive and limited to recording 1-2 cells. While extracellular multielectrode arrays can record multiple cells, they are pre-fabricated and thus can only probe fixed locations. Optical detection of electric activities provides the needed spatial flexibility. Calcium sensors such as GcaMP have a slow time response and not suitable to record fast-spiking pacemaker neurons such as dopaminergic neurons. Voltage-sensitive fluorescence proteins and dyes have much faster time response, but their recording time is usually limited by photobleaching. In this project, we will demonstrate an orthogonal approach of optical recording. This method, Electrochromic Optical Recording of Electric potentials (ECORE) makes use of a unique material property – optical absorption of an electrochromic film depends on applied voltages. We detect the optical reflection of an electrochromic film to read out cellular electrical activities. The method is truly label-free, i.e. free of any molecular probes that need to be incorporated into cells and perturb cellular physiology, and not limited by photobleaching or photo-toxicity. In preliminary work, we have built a sensitive optical setup that is able to detect the reflectivity change of the electrochromic film in response to electrical potentials as small as 10 microvolts. Indeed, we have used ECORE to successfully record single-cell action potentials in neurons, cardiomyocytes, and brain tissues. With this project, we plan to dramatically expand ECORE capabilities by developing a scanning ECORE platform for parallel detection and an ECORE microscope for subcellular measurement of neuroelectric activities. We will use ECORE to probe the functional connectivity of dopaminergic neurons in midbrain area. Accomplishment of this work will result in a new class of electrophysiological tools that can be used by other research groups.

项目摘要 要理解一个由相互连接的神经元组成的网络如何接收、存储和处理信息， 并行和高质量的神经电信号记录。细胞内记录技术，如贴片 钳是侵入性的，并且仅限于记录1-2个细胞。虽然细胞外多电极阵列可以记录多个 细胞，它们是预制的，因此只能探测固定的位置。电活动的光学检测 提供所需的空间灵活性。钙传感器如GcaMP具有缓慢的时间响应， 适合于记录快速尖峰起搏神经元，例如多巴胺能神经元。电压敏感 荧光蛋白和染料具有快得多的时间响应，但它们的记录时间通常受到以下因素的限制： 光漂白 在这个项目中，我们将展示一个正交的光学记录方法。这种方法，电致变色 电势的光学记录（ECORE）利用独特的材料性质-光学吸收 电致变色膜的厚度取决于所施加的电压。我们检测电致变色膜的光学反射 来读出细胞的电活动该方法是真正无标记的，即不需要任何分子探针 掺入细胞并扰乱细胞生理学，而不受光漂白或光毒性的限制。 在初步工作中，我们已经建立了一个敏感的光学装置，能够检测到的反射率变化的 电致变色膜响应小至10微伏的电势。我曾用过， 成功记录神经元、心肌细胞和脑组织中的单细胞动作电位。与此 项目，我们计划通过开发一个扫描ECORE平台， 平行检测和ECORE显微镜用于神经电活动的亚细胞测量。我们将 应用ECORE技术探讨中脑多巴胺能神经元的功能连接。完成 这项工作将产生一种新的电生理工具，可供其他研究小组使用。