Optimization, Application, and Dissemination of Imaging Modules for High-speed Mesoscopic Volumetric Recording of Neuroactivity in Scattering Brains

项目总结/摘要 最近的一些观察表明，哺乳动物大脑中复杂的脑功能来自高度并行的计算，其中有关感觉输入，内部状态和行为参数的信息被映射到高度分布的全脑神经元群体。这需要神经技术，允许在清醒和行为动物的生理时间尺度和细胞分辨率上大规模记录组织深度和大脑区域的神经活动。虽然基于双光子扫描荧光显微镜（2 p M）和遗传编码钙指示剂（GECIs）作为神经活动报告物的组合的光学工具开发的最新进展旨在通过开发更快、更大规模和体积的钙（Ca 2+）成像技术来解决这些需求，但在这种情况下，一个根本的未解决的挑战是在速度、分辨率和记录体积的大小。我们的实验室最近建立了这种最佳记录方案的标准，这导致了一种新的高速体积Ca 2+成像方法的实现，称为光珠显微镜（LBM）。通过LBM，我们已经证明了在清醒的行为小鼠的两个皮质半球内以高达100万个神经元的单细胞分辨率对神经活动进行荧光寿命有限的体积记录。在这个项目中，我们将采取多管齐下的策略，对我们的LBM技术进行优化，生物应用和有效传播，同时扩展其性能。这将使我们的LBM技术更强大、更简单、更用户友好。为了使其能够广泛有效地传播，在项目的第二部分，我们将利用α测试人员的反馈来设计、构建和传播我们系统的β原型，这些原型将分发给几个最终用户实验室，这些实验室将在他们的生物学问题中测试和应用我们的LBM技术。这个β原型也将成为我们技术商业传播的基础，同时也是开源传播的基础。

Project Summary / Abstract A number of recent observations suggest that complex brain functions in the mammalian brain emerge from highly parallel computation in which information about sensory inputs, internal states, and behavioral parameters are mapped onto highly distributed brain-wide neuronal populations. This calls for neurotechnologies that allow for large-scale recording of neuro-activity across tissue depths and brain regions at physiological timescales and cellular resolution in awake and behaving animals. While recent advancements in optical tool development based on the combination of two-photon scanning fluorescence microscopy (2p M) and genetically-encoded calcium indicators (GECIs) as reporters of neuro-activity have been aimed at addressing these needs by developing faster, larger-scale, and volumetric calcium (Ca2+) imaging technologies, a fundamental unsolved challenge in this context is navigating the inherent tradeoffs between speed, resolution, and the size of the recording volume in a principled and scalable manner. Our lab has recently established criteria for such optimal recording schemes which has led to the realization of a new high-speed volumetric Ca2+ imaging approach termed Light Beads Microscopy (LBM). Through LBM, we have demonstrated fluorescence lifetime limited volumetric recording of neuro-activity at a single-cell resolution of up to 1 million neurons within both cortical hemispheres of awake, behaving mice. In this project, we will pursue a multipronged strategy towards the optimization, biological applications, and effective dissemination of our LBM technology while extending its performance. This will result in a more robust, less complex, and more user-friendly version of our LBM technology. To enable its broad and effective dissemination, in the second part of the project, we will utilize feedback from our α-testers to design, build, and disseminate β-prototypes of our system that will be distributed to several end-user laboratories who will be testing and applying our LBM technology in the context of their biological questions. This β-prototype will also form the basis for commercial dissemination of our technology as well as a parallel effort for its open-source dissemination.