Efficient Two-Photon Voltage Imaging of Neuronal Populations at Behavioral Timescales

行为时间尺度神经元群的高效双光子电压成像

• 批准号: 10516906
• 负责人: Jerry L Chen
• 金额: $ 133.67万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10516906
• 关键词: Action Potentials; Address; Algorithms; Anatomy; Animal Behavior; Behavior; Behavioral; Biosensor; Brain; Brain Diseases; Budgets; Calcium; Communication; Crystallization; Data; Detection; Development; Electrophysiology (science); Engineering; Ensure; Fiber; Financial compensation; Fluorescence; Generations; Goals; Hot Spot; Hybrids; Image; Image Analysis; Label; Lasers; Learning; Length; Light; Mammalian Cell; Measurement; Measures; Methods; Microscope; Molecular; Monitor; Motion; Mus; Mutagenesis; Neocortex; Neurons; Neurosciences; Noise; Opsin; Optical Methods; Optics; Orthologous Gene; Photobleaching; Photons; Physiologic pulse; Play; Population; Process; Property; Protein Engineering; Reporting; Research Personnel; Resolution; Risk; Scanning; Scientist; Signal Transduction; Solid; Somatosensory Cortex; Speed; Supervision; System; Systems Analysis; Techniques; Testing; Time; Tissues; Variant; adaptive optics; analysis pipeline; base; cost effective; deep learning; denoising; design; experimental study; flexibility; genetic information; improved; in vivo; in vivo imaging; instrumentation; light scattering; multiplexed imaging; mutant; neuronal cell body; neuronal circuitry; new technology; novel; parallel computer; patch clamp; photonics; screening; sensor; spatiotemporal; tool; two-photon; voltage

PROJECT SUMMARY Understanding how information is processed in the mammalian neocortex has been a longstanding question in neuroscience. While the action potential is the fundamental bit of information, how these spikes encode representations and drive behavior remains unclear. In order to adequately address this problem, it has become apparent that experiments are needed in which activity from large numbers of neurons can be measured in a detailed and comprehensive manner across multiple timescales. Direct measurements of action potentials have primarily been achieved by electrophysiology. However, such measurements cannot easily be combined with other methods to assess the connectivity and molecular properties of neurons. Integrating functional, anatomical, and genetic information is critical for understanding how neuronal circuits are organized and computed. There have been long-standing efforts in developing optical methods for measuring neuronal activity due to its compatibility to simultaneously measure connectivity and molecular identity using fluorescent labeling techniques. We have developed a two-photon-excitable genetically-encoded voltage-sensitive indicator and ultra-fast two-photon microscope that enables optical measurements of action potentials deep into the brain. However, imaging at high signal-to-noise beyond several minutes remains challenging due to photo-bleaching and risks of photo- damage. In order for these new technologies to become more robust for neuroscience applications, it is necessary to improve upon the stability, reliability, and efficiency of two-photon voltage imaging. To achieve this, it requires a concerted effort between optical engineers, protein engineers, and computational scientists to optimize instrumentation, sensors, and image analysis for broad dissemination. This multi-investigator effort proposes to advance two-photon voltage imaging to enable sustained tracking of population activity at timescales of animal behavior and learning.

项目概要 了解哺乳动物新皮层如何处理信息一直是一个长期的问题 神经科学中的问题。虽然动作电位是信息的基本位，但这些信息如何 尖峰编码表征和驱动行为仍不清楚。为了充分解决这个问题 问题，很明显，需要进行实验，其中来自大量的活动 可以在多个时间尺度上以详细而全面的方式测量神经元。直接的 动作电位的测量主要通过电生理学来实现。然而，这样的 测量不能轻易地与其他方法结合来​​评估连通性和分子 神经元的特性。整合功能、解剖和遗传信息对于 了解神经元回路是如何组织和计算的。已经有长期的努力 开发用于测量神经元活动的光学方法，因为它同时兼容 使用荧光标记技术测量连接性和分子身份。我们开发了一个 双光子激发基因编码电压敏感指示器和超快双光子显微镜 这使得能够对大脑深处的动作电位进行光学测量。然而，在高成像 由于光漂白和光漂白的风险，超过几分钟的信噪比仍然具有挑战性。 损害。为了使这些新技术在神经科学应用中变得更加强大，需要 提高双光子电压成像的稳定性、可靠性和效率是必要的。到 要实现这一目标，需要光学工程师、蛋白质工程师和 计算科学家优化仪器、传感器和图像分析以实现广泛的应用 传播。这项多位研究人员的努力建议推进双光子电压成像，以实现 在动物行为和学习的时间尺度上持续跟踪种群活动。