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.

摘要 大脑研究的一个主要目标是对行为过程中神经元组的动力学进行成像。虽然 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，我们的工具是 在单个神经元水平上评估功能通常只允许进入大脑的小区域， 时间分辨率有限。活动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，非侵入性的测量，从单个神经元到完整的整个群体， 神经系统，并且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。 散射脑中的细胞分辨率成像通常通过多光子显微镜（MPM）实现。的 该提案的重点是开发、实施和推广新一代多光子成像工具 和基因编码的指标，允许深入，快速，大规模的结构和功能成像， 细胞和亚细胞分辨率。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馈送到源， 并将按需脉冲与显微镜扫描系统同步，AES将 将常规MPM转换为仅激发感兴趣区域的“随机访问”MPM。我们将整合AES 与高速扫描仪，导致新的AES-MPM，将提供> 10倍的成像速度提高，或 成像的神经元数量。我们将联合收割机将康奈尔大学的AES工作与的开发工作结合起来 基因编码电压指示器（GEVI）在斯坦福大学和Janelia研究校园，并证明了 AES-MPM在体内小鼠脑中成像GEVI标记的神经元。我们还将联合收割机与 多光子显微镜，以展示遗传编码Ca指示剂（GECI）的大规模成像。的 研究涉及PI（徐）、合作研究者Michael Lin（斯坦福大学）和Karel之间的密切互动 Svoboda（Janelia）.此外，调查人员将与工业合作伙伴密切合作，探索 将AES转化为商业可用的系统，这为广泛传播提供了直接途径。