C5: Optical Instrumentation

项目摘要/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 外设仪器，确保我们的研究人员拥有最好的可用技术。总体目标 这一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 神经活动的记录和扰动。这些方法依赖于光学仪器，例如 多光子显微镜、基于激光的光遗传微扰系统和宽视场显微镜。我们的 实验还依赖于实时生成和控制虚拟现实环境的设备 动物执行任务的机器。为了充分利用大脑倡议中的新技术，我们将 随着该领域的前沿向前发展，创新和升级仪器和方法。 核心的第一个目标是实现、支持和改进双光子和三光子 显微镜，用于钙成像，并与我们的老鼠行为设置集成在一起。我们会 保持四个中视场，双光子显微镜，以及我们的中视镜，它可以成像 以高有效率从两个或多个相隔很远的区域或层中分离出来。这个介镜是我们的核心 研究非相邻区域之间神经相互作用的仪器。我们还将在最近实施一项 开发的技术极大地增加了同时记录的神经元的数量。最后，我们会 维护和升级我们的三光子显微镜，用于皮质以下和深层区域的成像，以及 目的：提供血管成像技术，用于钙离子成像与电子显微镜的配准。 第二个目标是支持和改进广域成像仪器。我们将保持一个 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，并根据需要将其集成到新的行为系统中。这台仪器是用来绘制 在介观尺度上，通过钙成像观察视觉区域和神经活动的流动。 第三个目标将是实施、支持和改进我们的光遗传和成像组合 仪器仪表。我们建立了一台同时进行双光子成像和单细胞双光子成像的仪器 光刺激。此外，我们正在重新设计我们开发的一种结合了细胞分辨率的仪器 多细胞同时光发生微扰与神经细胞大尺度钙成像 种群，这将被用来研究动物的行为如何受到序列变化的影响 行为任务中的神经激活。总而言之，光学仪器核心将提供无缝 在所有研究项目中支持显微镜和虚拟现实实验。更广泛地说，我们预计 我们广泛使用的开创性和标准化的方法来可视化神经活动和 啮齿动物在虚拟现实中的实验将对神经科学界具有很高的价值。

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.