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成像一组神经元在行为过程中的动态。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 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，并且 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、非侵入性测量，范围从单个神经元到完整的整个种群 神经系统，并很容易与遗传方法相结合，提供特定细胞类型的记录。 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是通过多光子显微镜(MPM)实现的。这个 这项提案的重点是开发、实施和传播新一代多光子成像工具 以及基因编码的指示器，允许对结构和功能进行深度、快速和大规模的成像 细胞和亚细胞分辨率。为了达到“光子预算”所定义的基本极限，我们将 研制自适应激励源(AES)。通过将样品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，所述AES将 将传统的MPM转换为仅激发感兴趣区域的“随机访问”MPM。我们将集成AES 使用高速扫描仪，产生新的AES-MPM，它将使成像速度提高10倍或 被成像的神经元的数量。我们将把康奈尔大学的AES工作与 斯坦福大学和珍妮莉亚研究校园的遗传编码电压指示器(GEVI)，并展示了 应用AES-MPM对小鼠脑内GEVI标记神经元进行活体成像我们还将把AES与一个 多光子介镜显示遗传编码的钙指示剂(GECI)的大规模成像。这个 研究涉及派(徐)、合作调查员迈克尔·林(斯坦福大学)和卡雷尔之间的密切互动 斯沃博达(珍妮莉亚)。此外，调查人员将与行业合作伙伴密切合作，探索潜在的 将AES翻译成商业可用的系统，这提供了一条广泛传播的直接途径。