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 了解互连神经元网络如何接收，商店和流程信息需要 神经信号的平行和高质量记录。细胞内记录技术，例如补丁 夹具具有侵入性，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，因此只能探测固定位置。电动活动的光学检测 提供所需的空间灵活性。诸如GCAMP之类的钙传感器的时间响应缓慢而不是 适用于记录快速刺激的起搏器神经元，例如多巴胺能神经元。电压敏感 荧光蛋白和染料的时间响应更快，但它们的记录时间通常受到 光漂白。 在这个项目中，我们将展示一种正交记录的正交方法。这种方法，电致色素 电势的光学记录（ECORE）利用了独特的材料特性 - 光学滥用 电染料膜的含量取决于施加的电压。我们检测到电致色膜的光学反射 读取细胞电活动。该方法确实没有标签，即没有任何需要的分子问题 掺入细胞和扰动细胞生理学中，不受光漂白或光毒性的限制。 在初步工作中，我们建立了一个敏感的光学设置，能够检测到反射率的变化 响应小至10微伏的电势响应的电染料膜。确实，我们使用了生态 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生态平台来大大扩展生态功能 平行检测和用于神经电活动亚细胞测量的生态显微镜。我们将 使用Ecore探测中脑区域多巴胺能神经元的功能连通性。成就 这项工作将导致其他研究小组可以使用的新型电生理工具。