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: 10264522
• 负责人: BRIDGET Olivia CARRAGHER
• 金额: $ 78.7万
• 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/10264522
• 关键词: Biological; Cells; Cryoultramicrotomy; Data Collection; Ensure; Equipment; Fluorescence; Freezing; Funding; Goals; In Situ; Ions; Lifting; Light Microscope; Methods; Modernization; Preparation; Resolution; Sampling; Services; Specimen; Time; Training; Training Programs; Visualization; career; design; instrumentation; light microscopy; method development; pressure; screening; skills; tomography

Summary The primary goal of the National Center for In-situ Tomographic Ultramicroscopy (NCITU) is to provide user access and service towards preparing specimens for direct visualization of macromolecular associations and organization, particularly in native cells. NCITU is 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. A highly qualified technical team provides 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. As NCITU develops, it 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. This is a request for supplemental funding to purchase instrumentation that will expand and enhance the capabilities of the center. We are requesting: (i) a correlative fluorescence light microscope integrated into the FIB/SEM; (ii) a modern high pressure freezer; and (iii) a dedicated cryo fluorescence confocal light microscope.

概括 国家原位断层扫描超显镜（NCITU）的主要目标是提供 用户访问和服务用于准备标本以直接可视化大分子 关联和组织，特别是在本地细胞中。 NCITU致力于发展和 提供对原位冷冻标本准备的标准方法和高级方法的访问，以及 通过一个旨在为广泛服务的冷冻交叉训练计划传播这一专业知识 各种技能水平和职业目标。一个高素质的技术团队提供直接动手 服务，指导和协助，以确保准备最高的样品和网格 质量，最短的时间。随着NCITU的发展，它将为 层析成像的原位样品制备包括：培养生物样品，陷入冻结和 高压冻结，冷冻显微镜（冰冻），冷冻粉状切开术，聚焦离子束铣削 （纤维填充），包括提出和初始的冷冻仪筛选，然后提交标本 高分辨率冷冻数据收集的网络中心。 这是购买仪器的补充资金的要求，以扩大和增强 中心的功能。我们要求：（i）相关荧光光显微镜 集成到FIB/SEM中； （ii）现代的高压冰柜； （iii）专用的冷冻荧光 共聚焦光显微镜。