Label-free Optical Recording of Neuroelectric Activities

神经电活动的无标记光学记录

基本信息

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
项目类别：
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

项目摘要

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.

项目概要了解互连神经元网络如何接收、存储和处理信息需要细胞内记录技术（例如贴片）的并行和高质量记录。钳是侵入性的，只能记录 1-2 个细胞，而细胞外多电极阵列可以记录多个细胞。细胞，它们是预制的，因此只能探测固定位置的电活动。提供所需的空间灵活性，例如 GcaMP 的时间响应较慢。适合记录快速尖峰起搏神经元，例如多巴胺能神经元。荧光蛋白和染料具有更快的时间响应，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，我们将演示光学记录的正交方法。电势光学记录 (ECORE) 利用独特的材料特性 - 光吸收电致变色薄膜的光学反射取决于所施加的电压。该方法真正无需标记，即无需任何分子探针。被纳入细胞并扰乱细胞生理学，并且不受光漂白或光毒性的限制。在前期工作中，我们建立了一个灵敏的光学装置，能够检测物体的反射率变化响应小至 10 微伏电势的电致变色薄膜事实上，我们使用了 ECORE。成功记录神经元、心肌细胞和脑组织的单细胞动作电位。项目中，我们计划通过开发扫描 ECORE 平台来大幅扩展 ECORE 功能我们将使用并行检测和 ECORE 显微镜来测量神经电活动的亚细胞。使用 ECORE 来探测中脑区域多巴胺能神经元的功能连接。这项工作将产生一类可供其他研究小组使用的新型电生理学工具。