Expanding Access to Planar Illumination Microscopy in a Neuroimaging Core

描述(由申请人提供)：为了支持NINDS任务的追求，我们建议向华盛顿大学神经科学界介绍一种高影响力的成像新技术。我们打算提供核心仪器来执行平面照明显微镜(也称为光片显微镜)，以催化新的成像努力，旨在监测大规模神经元集合。最近联邦政府宣布的大脑活动地图的目标是神经生理学，以期在未来十年实现快速增长和发展。高通量神经生理学最有前途的技术之一是使用光片的荧光成像。我们的实现，物镜耦合平面照明(OCPI)显微镜，通过同时照射物镜焦面中的所有像素，实现了比双光子显微镜快数百或数千倍的成像速度，从而消除了通过一次扫描一个像素来收集图像的需要。该方法实现了快速成像，同时具有高灵敏度和低光毒性，因此特别适合于长期记录周期。定制的OCPI仪器将是现有成像中心--贝克韦尔神经成像设施的核心产品。因此，本提案描述了引入新的OCPI显微镜的计划和时间表，以及我们将建立的监督，以确保公平和广泛地使用它。我们还强调，我们在这些基于社区的努力方面拥有最近的经验：我们成功地引入了该仪器的早期版本作为核心设施，现在可以预见更多的神经科学家社区需要更多的更快、更灵活的版本。我们计划为OCPI用户提供设施支持，包括一名提供维护和培训的设施经理、一个用于图像分析的数据中心和一名帮助进行定制数据分析的数据经理。我们提供了一系列可量化的里程碑，用来评估在提供这种新的高影响力技术方面的进展。到四年结束时，我们预计吴神经科学社区至少有20名不同用户的稳定基础，使用率达到或接近全职。

DESCRIPTION (provided by applicant): To support pursuit of the NINDS mission, here we propose to introduce a high-impact novel technology for Imaging to the Washington University Neuroscience community. We intend to provide Core Instrumentation to perform Planar Illumination Microscopy (also known as light sheet microscopy) to catalyze new imaging efforts that aim to monitor large-scale neuronal ensembles. The recent Federal announcement of the Brain Activity Map targets neurophysiology for rapid growth and development in the coming decade. One of the most promising techniques for high-throughput neurophysiology is fluorescence imaging using light sheets. Our implementation, Objective-Coupled Planar Illumination (OCPI) microscopy, achieves imaging speeds that are hundreds or thousands of times faster than two-photon microscopy by simultaneously illuminating all pixels in the objective's focal plane, thereby eliminating the need to collect images by scanning one pixel at a time. The method achieves fast imaging simultaneously with high sensitivity and low phototoxicity and is therefore particularly well-suited to long-term recording periods. The custom-built OCPI instrument will be a Core offering within an existing Imaging Center, the Bakewell Neurolmaging Facility. Therefore, this proposal describes a plan and a schedule by which the new OCPI microscope will be introduced, and the oversight we will establish to ensure that its use is made available fairly and broadly. We also emphasize that we have recent experience in precisely these community-based efforts: we successfully introduced earlier version of this instrumentation as a Core facility and now can foresee the need for more access by a larger community of Neuroscientists to a faster and more flexible version. We plan Facility support for OCPI users that includes a facility manager to provide maintenance and training, a data center for image analysis, and a data manager to help with customized data analysis. We provide a series of quantifiable milestones by which to evaluate progress in offering this new high-impact technology.' By the end of the 4-yr period, we anticipate a steady base of at least 20 different users from across the diverse WU Neuroscience community, and a usage rate at or near full-time.