Optimization, application and dissemination of high-speed hybrid multiphoton volumetric imaging technologies

高速混合多光子体积成像技术的优化、应用和推广

• 批准号: 9921918
• 负责人: Attila Losonczy
• 金额: $ 161.62万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9921918
• 关键词: Anatomy; Area; Behavioral; Biological; Brain; Brain region; Calcium; Cells; Characteristics; Cognitive; Collaborations; Communities; Development; Dissection; Dorsal; Engineering; Feedback; Functional Imaging; Goals; Head; Hippocampus (Brain); Hybrids; Image; Imaging technology; Industrialization; Industry; Joints; Learning; Light; Microscopy; Modeling; Mus; Neurobiology; Neurons; Neurosciences; Optics; Pattern; Performance; Population; Population Dynamics; Resolution; Scanning; Sensory; Speed; System; Technology; Testing; Time; Tissues; Universities; awake; base; cell type; commercialization; cost; dentate gyrus; design; flexibility; foot; imaging modality; imaging platform; in vivo; insight; light microscopy; motor behavior; neural circuit; neuronal patterning; open source; photonics; prototype; spatiotemporal; temporal measurement; three photon microscopy; tool; two-photon; way finding

PROJECT SUMMARY / ABSTRACT Understanding how cognitively-relevant behavioral functions emerge from activity patterns of identified cell- types is predicated on the ability to record large-scale ensemble dynamics from genetically-identified and longitudinally-tracked neuronal populations across multiple brain regions and layers with high spatial and temporal resolution over behaviorally-relevant time-scales. Two-photon scanning microscopy in combination with genetically-encoded calcium (Ca2+) indicators is currently the most essential tool for in vivo optical recording of neuronal activity, its application to deep brain regions. However, currently the commercially available 2pM systems are limited in their applications due to constraints related to the obtainable imaging depth, volumetric field-of-view (VFOV), and temporal resolution at which neuronal population dynamics can be effectively captured. We have recently developed and demonstrated the proof of principle of a new high-speed volumetric Ca2+-imaging platform termed Hybrid Multiplexed Sculpted Light (HyMS) Microscopy that combines 2pM with three-photon microscopy (3pM). HyMS allows for volumetric recording of neuroactivity at single-cell resolution within volumes up to ~1 × 1 × 1.22 mm at up to 17 Hz in cortical as well as sub-cortical regions of awake behaving mice. The impact of this tool will depend on a successful optimization, neurobiological application and dissemination strategy within the neuroscience community. While we will provide open source access for technically skilled labs, given the technical complexity and costs of such a system, the most effective strategy is through partnership with industry and through commercialization of the system. Here we propose a roadmap towards this objective. Building on our current existing system, we will implement a number of technical refinements and optimizations. Leveraging the ongoing collaboration with the Losonczy Lab at the Columbia University, we will use our optimized HyMS system to perform high-speed multiphoton volumetric Ca2+ imaging of functional circuitry across the entire depth of the mouse dorsal hippocampus (HPC), encompassing all major regions of the HPC trisynaptic circuitry. This application will provide us valuable feedback for further optimization and refinement and development of our HyMS prototype system. In parallel, we will develop together with our industrial partner a first prototype of the HyMS system (-HyMS) This prototype will be again used and tested by the Losonczy Lab. The obtained insights and user feedback from their application will drive the development of a beta prototype (-HyMS) which will be used to engage broader local users as beta testers. 9 user labs, mainly from the NYC area, with a broad range of biological questions and applications, will participate as beta testers and provide us with iterative user feedback which will ultimately drive and be incorporated both into the into the commercialization of HyMS as well its open source model of the access to this technology.

项目总结/摘要 了解认知相关的行为功能是如何从已识别细胞的活动模式中产生的， 类型是基于从基因鉴定和 跨多个大脑区域和层的神经元群体具有高空间和 行为相关时间尺度上的时间分辨率。双光子扫描显微镜的组合 与遗传编码的钙（Ca 2+）指标是目前最重要的工具，在体内光学 神经元活动的记录及其在脑深部区域的应用然而，目前商业 由于与可获得的成像相关的约束，可用的2pM系统在其应用中受到限制 深度、体积视场（VFOV）和时间分辨率，在这些分辨率下，神经元群体动力学可以被 有效捕获。我们最近开发并证明了一种新的高速 体积Ca 2+成像平台，称为混合多路雕刻光（HyMS）显微镜， 2pM，三光子显微镜（3 pM）。HyMS允许在单个细胞上的神经活动的体积记录 在皮质和皮质下区域，分辨率高达约1 × 1 × 1.22 mm，频率高达17 Hz， 清醒的老鼠该工具的影响将取决于一个成功的优化，神经生物学 在神经科学界的应用和传播策略。虽然我们将提供开源 技术熟练的实验室，考虑到这种系统的技术复杂性和成本， 战略是通过与工业界的伙伴关系和通过系统的商业化。在这里，我们提出一个 实现这一目标的路线图。在现有制度的基础上，我们会推行多项 技术改进和优化。利用与Losonczy实验室在 哥伦比亚大学，我们将使用我们优化的HyMS系统进行高速多光子体积 小鼠背海马（HPC）整个深度的功能回路的Ca 2+成像， 包括HPC三突触电路的所有主要区域。这个应用程序将为我们提供宝贵的 我们的HyMS原型系统的进一步优化和完善和发展的反馈。同时， 我们将与我们的工业合作伙伴共同开发HyMS系统的第一个原型（HyMS-HyMS）。 Losonczy实验室将再次使用原型并进行测试。获得的见解和用户反馈，从他们的 应用程序将推动开发一个测试版原型（EQ-HyMS），该原型将用于吸引更广泛的当地 用户作为Beta测试人员。9个用户实验室，主要来自纽约地区，有广泛的生物学问题， 应用程序，将作为beta测试人员参与，并为我们提供迭代的用户反馈，最终将 推动并融入HyMS的商业化以及其开源模型， 获得这项技术。