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

总结 国家原位层析超显微镜中心（NCITU）的主要目标是提供 为制备用于大分子直接可视化的样品的用户访问和服务 联系和组织，特别是在原生细胞中。NCITU致力于开发和 提供原位冷冻ET标本制备的标准和先进方法，以及 通过冷冻ET交叉培训计划传播这一专业知识， 各种技能水平和职业目标。高素质的技术团队提供直接的实践操作 服务、指导和帮助，以确保样品和网格的制备具有最高的 质量和在最短的时间内。随着NCITU的发展，它将提供全方位的服务， 用于断层扫描的原位样品制备，包括：培养生物样品，柱塞冷冻， 高压冷冻，冷冻光显微镜（cryoLM），冷冻超微切片，聚焦离子束铣削 （FIB研磨），包括提出，以及在将标本提交给 用于高分辨率cryoET数据收集的网络集线器。 这是一个补充资金的要求，购买仪器，将扩大和加强 中心的能力。我们要求：（一）相关荧光显微镜 集成到FIB/SEM中;（ii）现代高压冷冻机;以及（iii）专用的低温荧光 共聚焦光学显微镜

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.