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: 10471831
• 负责人: Attila Losonczy
• 金额: $ 95.45万
• 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/10471831
• 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; dissemination strategy; 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.

项目摘要 /摘要 了解与认知相关的行为函数如何从已识别的细胞的活性模式中出现 预测类型是关于从遗传识别和 多个大脑区域和高空间和层的纵向跟踪神经元种群 与行为相关的时间尺度上的临时分辨率。组合的两光子扫描显微镜 具有一般编码的钙（CA2+）指标目前是体内光学的最重要的工具 记录神经元活性，其应用于深脑区域。但是，目前在商业上 由于与可获得成像有关的限制，可用的2pm系统在其应用中受到限制 深度，体积视野（VFOV）和临时分辨率，即神经元人口动态的分辨率 有效捕获。我们最近开发并证明了新的高速原理证明 体积Ca2+成像平台称为混合雕刻光（HYMS）显微镜 2pm，三光子显微镜（下午3点）。赞美诗允许在单细胞时进行大量记录神经活性 在皮质和皮质下最多17 Hz的卷内分辨率高达〜1×1×1.22 mm。 唤醒行为的老鼠。该工具的影响将取决于成功的优化，神经生物学 神经科学界的应用和传播策略。虽然我们将提供开源 鉴于这种系统的技术复杂性和成本，最有效的技术复杂性和成本，访问技术熟练的实验室 策略是通过与行业合作和系统的商业化来进行的。在这里我们提出一个 通往这个目标的路线图。在当前现有系统的基础上，我们将实施许多 技术改进和优化。利用与Losonczy实验室的持续合作 哥伦比亚大学，我们将使用优化的赞美诗系统来执行高速多光体积 跨小鼠背海马（HPC）整个深度的功能电路的CA2+成像 涵盖HPC三突触电路的所有主要区域。该申请将为我们提供宝贵的 反馈，以进一步优化，完善和开发我们的赞美诗原型系统。并联， 我们将与我们的工业合作伙伴一起发展一个赞美诗系统（-hyms）的第一个原型 原型将由Losonczy Lab再次使用和测试。从其获得的见解和用户反馈 应用将推动开发Beta原型（-HYMS），该原型将用于吸引更广泛的本地 用户作为Beta测试人员。 9个用户实验室，主要来自纽约市区，有各种各样的生物学问题和 应用程序将作为Beta测试人员参与，并为我们提供迭代用户反馈，最终将 驱动器并将两者都纳入赞美诗的商业化以及其开源模型 访问这项技术。