Expanding Access to Planar Illumination Microscopy in a Neuroimaging Core

扩大神经影像核心中平面照明显微镜的使用范围

• 批准号: 8662909
• 负责人: Paul H Taghert
• 金额: $ 30.36万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8662909
• 关键词: Address; Ally; Bioinformatics; Brain; Communities; Core Facility; Coupled; Custom; Data; Data Analyses; Elements; Ensure; Growth and Development function; Housing; Image; Image Analysis; Imaging technology; Laboratories; Light; Lighting; Maintenance; Maps; Methods; Microscope; Microscopy; Mission; Monitor; National Institute of Neurological Disorders and Stroke; Neurons; Neurosciences; Neurosciences Research; Optics; Pharmacology; Phototoxicity; Physiology; Research; Research Activity; Scanning; Schedule; Series; Specialist; Speed; System; Techniques; Technology; Time; Training; Universities; Washington; base; design; experience; flexibility; fluorescence imaging; high throughput technology; imaging modality; innovation; instrument; instrumentation; nervous system disorder; neuroimaging; neurophysiology; new technology; novel; public health relevance; rapid growth; two-photon

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.

描述（由申请人提供）：为了支持NINDS使命的追求，在这里，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），以促进旨在监测大规模神经元集合的新成像工作。最近联邦公布的脑活动地图的目标是神经生理学的快速增长和发展，在未来十年。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，双光子耦合平面照明（OCPI）显微镜，实现了数百或数千倍的成像速度比双光子显微镜，同时照亮物镜的焦平面中的所有像素，从而消除了需要通过扫描一个像素的时间来收集图像。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期。定制的OCPI仪器将成为现有成像中心贝克韦尔神经成像设施的核心产品。因此，本提案描述了一个计划和时间表，通过该计划和时间表，将引入新的OCPI显微镜，以及我们将建立的监督，以确保其公平和广泛的使用。我们还强调，我们最近在这些基于社区的努力中取得了经验：我们成功地将该仪器的早期版本作为核心设施引入，现在可以预见更大的神经科学家社区需要更多的访问更快，更灵活的版本。我们计划为OCPI用户提供设施支持，其中包括提供维护和培训的设施经理，用于图像分析的数据中心以及帮助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，以评估提供这种新的高影响力技术的进展。“到4年期结束时，我们预计来自不同WU神经科学社区的至少20个不同用户的稳定基础，以及全职或接近全职的使用率。