Close-loop, spatially addressable multiphoton functional imaging

闭环、空间可寻址多光子功能成像

• 批准号: 10246271
• 负责人: CHRIS XU
• 金额: $ 60.95万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10246271
• 关键词: 3-Dimensional; Amplifiers; Behavior; Biological; Brain; Brain region; Budgets; Calcium; Development; Functional Imaging; Generations; Genetic; Goals; Image; Imaging Device; Individual; Industrialization; Label; Measurement; Methods; Microscope; Monitor; Mus; Nervous system structure; Neurons; Neurosciences; Noise; Optics; Output; Performance; Photons; Physiologic pulse; Population; Research; Research Personnel; Resolution; Resource Sharing; Sampling; Scanning; Seeds; Signal Transduction; Source; Specimen; Speed; Structure; System; Techniques; Testing; Three-Dimensional Imaging; Time; TimeLine; Translating; Viral; Work; brain research; brain tissue; calcium indicator; cell type; design; experimental study; feeding; improved; in vivo; interest; multiphoton imaging; multiphoton microscopy; optical imaging; programs; relating to nervous system; temporal measurement; tool; transmission process; voltage

Abstract A major goal of brain research is to image the dynamics of groups of neurons during behavior. Although even the simplest behaviors involve interactions across multiple parts of the nervous system, our tools for assessing function at the level of individual neurons usually allow only access to small regions of the brain, and with limited temporal resolution. Optical recordings of activity are critical to probe neural systems because they provide high-resolution, non-invasive measurements, ranging from single neurons to entire populations in intact nervous systems, and are readily combined with genetic methods to provide cell type-specific recordings. Nevertheless, the limited spatial scale and temporal resolution remain a major challenge for optical imaging. Cellular-resolution imaging in scattering brains is typically achieved with multiphoton microscopy (MPM). The focus of this proposal is to develop, implement, and disseminate a new generation of multiphoton imaging tools and genetically encoded indicators that allow deep, fast, and large-scale imaging of structure and function with cellular and subcellular resolution. To approach fundamental limits defined by the 'photon budget', we will develop an adaptive excitation source (AES). By feeding the structural information of the sample to the source, and synchronizing the on-demand pulses with the microscope scanning system, the AES transforms a conventional MPM into a “random-access” MPM that only excites regions of interest. We will integrate the AES with high speed scanners, resulting in a new AES-MPM that will provide >10x improvement in imaging speed or the number of neurons imaged. We will combine the AES effort at Cornell with the development effort of genetically encoded voltage indicator (GEVI) at Stanford and Janelia Research Campus, and demonstrate the AES-MPM in imaging GEVI labeled neurons in mouse brains in vivo. We will also combine the AES with a multiphoton mesoscope to demonstrate large scale imaging of genetically encoded Ca indicator (GECI). The research involves close interactions between the PI (Xu), Co-investigator Michael Lin (Stanford), and Karel Svoboda (Janelia). Furthermore, the investigators will work closely with industrial partners to explore the potential of translating the AES into a commercially available system, which provides a direct path to broad dissemination.

抽象的 大脑研究的一个主要目标是对行为过程中神经元群的动态进行成像。 即使是最简单的行为也涉及神经系统多个部分的相互作用，这是我们的工具 在单个神经元水平上评估功能通常只允许访问大脑的一小部分区域，并且 时间分辨率有限的光学记录对于探测神经系统至关重要，因为它们 提供高分辨率、非侵入性测量，范围从单个神经元到完整的整个群体 神经系统，并且很容易与遗传方法结合以提供细胞类型特异性记录。 然而，有限的空间尺度和时间分辨率仍然是光学成像的主要挑战。 散射大脑中的细胞分辨率成像通常是通过多光子显微镜（MPM）实现的。 该提案的重点是开发、实施和传播新一代多光子成像工具 和基因编码的指示剂，可以对结构和功能进行深度、快速和大规模的成像 为了接近“光子预算”定义的基本极限，我们将 开发自适应激励源 (AES) 通过将样本的结构信息提供给源， 并将按需脉冲与显微镜扫描系统同步，AES 将 传统的 MPM 转变为仅激发感兴趣区域的“随机访问”MPM 我们将集成 AES。 与高速扫描仪，产生新的 AES-MPM，将提供超过 10 倍的成像速度或 我们将把康奈尔大学的 AES 工作与开发工作结合起来。 斯坦福大学和珍妮莉亚研究园的基因编码电压指示器（GEVI），并展示了 AES-MPM 在小鼠大脑中对 GEVI 标记的神经元进行体内成像 我们还将 AES 与 多光子介观镜展示了基因编码 Ca 指示剂 (GECI) 的大规模成像。 研究涉及 PI（Xu）、联合研究员 Michael Lin（斯坦福大学）和 Karel 之间的密切互动 此外，研究人员将与工业合作伙伴密切合作，探索潜力。 将 AES 转化为商用系统，为广泛传播提供了直接途径。