C5: Optical Instrumentation

项目概要/摘要：核心5，光学仪器 光学仪器核心将开发，实施，支持，集成和改进光学和 外围仪器，确保我们的研究人员拥有最好的技术。总体目标 这项U19合作的目的是阐明工作记忆和决策是如何通过互动来支持的。 神经元和大脑区域。为了实现这一目标，我们的研究项目需要大规模的方法， 记录和干扰神经活动。这些方法依赖于光学仪器，例如 多光子显微镜、基于激光的光遗传学扰动系统和宽视场显微镜。我们 实验还依赖于在真实的时间内生成和控制虚拟现实环境的设备， 动物们执行任务的方式。为了充分利用BRAIN计划中的新技术，我们将 创新和升级仪器和方法，作为该领域的前沿前进。 核心的第一个目标将是实施，支持和改进双光子和三光子 显微镜，用于钙成像，并与我们的小鼠行为设置集成。我们将 保持四个中等视野，双光子显微镜，沿着与我们的显微镜，可以成像 从两个或多个分开较远的区域或层以高的有效速率分离。这个显微镜是我们的核心 用于研究非相邻区域之间的神经相互作用的仪器。我们最近还将实施一项 开发的技术，大大增加了同时记录的神经元数量。最后我们将 维护和升级我们的三光子显微镜，用于大脑皮层以下和深层区域的成像， 以提供脉管系统成像，用于用电子显微镜记录钙成像。 第二个目标是支持和改进宽场成像仪器。我们会保持 基于背靠背物镜系统的定制宽视场显微镜，用于我们的鼠标虚拟现实 系统，并根据需要将其整合到新的行为系统中。该仪器用于绘制 视觉领域和流动的神经活动通过钙成像在介观尺度。 第三个目标将是实施，支持和改进我们的组合光遗传学和成像 仪器仪表我们建立了一个仪器，同时双光子成像和单细胞，双光子 光刺激此外，我们正在重新设计我们开发的一种仪器， 多个细胞的光遗传学扰动同时与神经细胞的大规模钙成像 种群，这将被用来研究动物的行为是如何受到影响的序列的变化， 行为任务中的神经激活总之，光学仪器核心将提供无缝的 支持所有研究项目中的显微镜和虚拟现实实验。更广泛地说，我们预计 我们的开创性和标准化方法的广泛可用性， 虚拟现实中的啮齿动物实验对神经科学界来说将是非常有价值的。

Project Summary/Abstract: Core 5, Optical Instrumentation The Optical Instrumentation Core will develop, implement, support, integrate, and improve optical and peripheral instrumentation, ensuring that our researchers have the best available technology. The overall aim of this U19 collaboration is to elucidate how working memory and decision-making are supported by interacting neurons and brain regions. To achieve this goal, our research projects require methods for large-scale recording and perturbation of neural activity. These methods depend on optical instrumentation, such as multi-photon microscopes, laser-based optogenetic perturbation systems, and widefield microscopy. Our experiments also rely on equipment that generates and controls, in real time, the virtual-reality environment in which the animals perform tasks. To take full advantage of the new technologies in the BRAIN Initiative, we will innovate and upgrade instrumentation and methods as the cutting edge of the field moves forward. The core’s first aim will be to implement, support, and improve two-photon and three-photon microscopes, which are used for calcium imaging and integrated with our mouse behavioral setups. We will maintain four mid-range field of view, two-photon microscopes, along with our mesoscope, which can image from two or more widely separated regions or layers at a high effective rate. This mesoscope is our core instrument for studying neural interactions between non-adjacent regions. We will also implement a recently developed technique that greatly increases the number of neurons simultaneously recorded. Finally, we will maintain and upgrade our three-photon microscope, used to image below cortex and in deep areas, and also to provide vasculature imaging for registering calcium imaging with electron microscopy. The second aim will be to support and improve widefield-imaging instrumentation. We will maintain a custom widefield microscope based on a back-to-back objective system for use with our mouse virtual-reality systems and integrate it into new behavioral systems as needed. This instrument is used to map boundaries of visual areas and the flow of neural activity via calcium imaging at the mesoscopic scale. The third aim will be to implement, support, and improve our combined optogenetic and imaging instrumentation. We built an instrument for simultaneous two-photon imaging and single-cell, two-photon photostimulation. In addition, we are redesigning an instrument we developed that combines cellular-resolution optogenetic perturbation of multiple cells simultaneously with large-scale calcium imaging of neural populations, which will be used to study how an animal's behavior is affected by changes in the sequences of neural activation during behavioral tasks. In summary, the Optical Instrumentation Core will provide seamless support for microscopy and virtual-reality experiments within all the research projects. More broadly, we expect that widespread availability of our pioneering and standardized methods for visualizing neural activity and for rodent experiments in virtual reality will be highly valuable to the neuroscience community.