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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要了解认知相关的行为功能如何从已识别细胞的活动模式中产生类型取决于从基因识别和记录大规模系综动态的能力跨多个大脑区域和层的纵向跟踪神经群体，具有高空间和行为相关时间尺度上的时间分辨率组合。具有基因编码的钙（Ca2+）指示剂是目前体内光学最重要的工具神经活动的记录及其在大脑深部区域的应用然而，目前商业化。由于与可获得的成像相关的限制，可用的 2pM 系统的应用受到限制深度、体积视场 (VFOV) 和时间分辨率，神经群体动态可以在这些分辨率下进行我们最近开发并演示了一种新的高速原理的证明。体积 Ca2+ 成像平台，称为混合多重雕刻光 (HyMS) 显微镜，结合了 2pM 与三光子显微镜 (3pM) 可以对单细胞的神经活动进行体积记录。在高达 17 Hz 的皮质和皮质下区域，体积内的分辨率高达 ~1 × 1 × 1.22 mm 该工具对清醒行为的小鼠的影响将取决于神经生物学的成功优化。神经科学界的应用和传播策略，同时我们将提供开源。考虑到此类系统的技术复杂性和成本，技术熟练的实验室的访问权限是最有效的战略是通过与工业界的合作以及系统的商业化来实现。为实现这一目标，我们将在现有系统的基础上实施一系列路线图。利用与Losonczy 实验室的持续合作。哥伦比亚大学，我们将使用我们优化的 HyMS 系统来执行高速多光子体积测量对小鼠背侧海马 (HPC) 整个深度的功能电路进行 Ca2+ 成像，涵盖 HPC 三突触电路的所有主要区域该应用将为我们提供有价值的信息。同时，进一步优化、完善和开发我们的 HyMS 原型系统。我们将与我们的工业合作伙伴一起开发 HyMS 系统的第一个原型 (-HyMS) Losonczy 实验室将再次使用和测试原型，并从他们获得的见解和用户反馈中进行测试。应用程序将推动测试原型 (-HyMS) 的开发，该原型将用于吸引更广泛的本地参与用户作为 beta 测试人员。 9 个用户实验室，主要来自纽约地区，具有广泛的生物学问题和应用程序，将作为 Beta 测试人员参与并向我们提供迭代的用户反馈，最终将推动并纳入 HyMS 及其开源模型的商业化获得这项技术。