The National Center for In-situ Tomographic Ultramicroscopy (NCITU)

国家原位断层超显微术中心 (NCITU)

• 批准号: 10264061
• 负责人: BRIDGET Olivia CARRAGHER
• 金额: $ 170万
• 依托单位: NEW YORK STRUCTURAL BIOLOGY CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10264061
• 关键词: Area; Biological; Cell Culture Techniques; Cells; Computers and Advanced Instrumentation; Cryoelectron Microscopy; Cryoultramicrotomy; Data Collection; Development; Electron Microscopy; Ensure; Environment; Equipment; Freezing; Goals; In Situ; Intention; Ions; Laboratory Research; Lifting; Methods; New York; Outcome; Preparation; Procedures; Protocols documentation; Research Personnel; Research Project Grants; Resolution; Running; Sampling; Science; Services; Specimen; Supervision; Techniques; Technology; Time; Tissues; Training; Training Programs; Visualization; Work; base; career; design; expectation; experience; improved; innovation; instrumentation; light microscopy; method development; new technology; novel; novel strategies; operation; pressure; screening; skills; structural biology; technology development; technology validation; tomography

Summary We propose to establish the National Center for In-situ Tomographic Ultramicroscopy (NCITU) to be based at the Simons Electron Microscopy Center (SEMC), which is a part of the New York Structural Biology Center (NYSBC), based in New York, NY. NCITU will be dedicated to developing and providing access to both standard and advanced methods for in-situ cryoET specimen preparation, and to disseminating this expertise through a cryoET cross-training program designed to serve a wide variety of skill levels and career goals. The primary goal of NCITU will be to provide user access and service towards preparing specimen for direct visualization of macromolecular associations and organization, particularly in native cells. A highly qualified technical team will provide direct hands-on service, guidance, and assistance to ensure that samples and grids are prepared with the highest quality and in the shortest time possible. An already well established cross-training program will be expanded and extended to provide training in every aspect of the cryoET specimen preparation pipeline. The operations at NCITU will build on already established practices at SEMC/NYSBC and benefit from the overall vibrant environment as well as the expertise of the highly trained staff already in place. NCITU will provide a full range of services for in-situ sample preparation for tomography including: culturing biological samples, plunge freezing and high pressure freezing, cryo-light microscopy (cryoLM), cryo- ultramicrotomy, focused ion beam milling (FIB-milling) including lift-out, and initial cryoTEM screening prior to submission of specimens to the network Hub for high-resolution cryoET data collection. We have already established workflows for many of these activities and have active research projects focused on expanding capabilities for automating and streamlining the entire workflow. We will continue to stay at the forefront of new developments in this area and we will incorporate new methods, technologies and workflows into the standard NCITU operating procedures as soon as these are stable and robust. While the goals of NCITU are not innately innovative, it is our intention to set the standard as one of the first world class, nationally accessible cryoET sample preparation facilities, to provide access and training that is freely and fairly available to all researchers in the US, and to continually strive to improve methods and enable science of the highest quality and value.

概括 我们建议建立国家原位层析成像超显镜（NCITU）的国家中心 Simons电子显微镜中心（SEMC），它是纽约结构生物学中心的一部分 （NYSBC），总部位于纽约，纽约。 NCITU将致力于开发和提供对这两个标准的访问 以及原位冷冻标本准备的高级方法，并通过A传播此专业知识 冷冻交叉训练计划旨在服务于各种技能水平和职业目标。 NCITU的主要目标是提供用户访问和服务以准备直接标本 大分子关联和组织的可视化，特别是在天然细胞中。高度合格 技术团队将提供直接的动手服务，指导和帮助，以确保样品和网格 以最高质量和最短的时间制备。已经建立了良好的交叉训练 计划将进行扩展和扩展，以在冷冻标本准备的各个方面提供培训 管道。 NCITU的运营将基于SEMC/NYSBC已经建立的实践，并从中受益 整体充满活力的环境以及训练有素的员工的专业知识已经到位。 NCITU将为断层扫描提供全方位的服务，以进行层析成像，包括：培养 生物样品，陷入冻结和高压冻结，冷冻显微镜（冰冻），冷冻 超微切开术，聚焦离子束铣削（纤维铣削），包括拔出以及初始的冷冻仪筛选 将标本提交给网络中心，以进行高分辨率冷冻数据收集。我们已经 为许多这些活动建立了工作流程，并拥有专注于扩展的积极研究项目 自动化和简化整个工作流程的功能。我们将继续保持新的最前沿 该领域的发展，我们将将新的方法，技术和工作流程纳入标准 NCITU操作程序立即稳定且健壮。 尽管NCITU的目标不是天生的创新性，但我们打算将标准设定为第一个标准之一 世界一流，全国可访问的冷冻样品准备设施，以提供访问和培训 美国的所有研究人员都可以自由且公平地使用，并不断努力改善方法并启用 最高质量和价值的科学。