Acquisition of 200kV Glacios Cryo Transmission Electron Microscope

项目摘要/摘要 我们计划在凯斯韦斯特购买最先进的200千伏冰川冷冻透射电子显微镜 后备大学医学院(CWRU SOM) 装配。低温电子显微镜(Cryo-EM)已经成为一种无与伦比的高分辨率工具 大分子的结构测定，大到40 kDa，大到几个丙二醛。 这项技术允许在溶液中、自然膜双层甚至其生理状态下对样品进行成像 蜂窝环境。这台新显微镜将扩展CWRU的冷冻-EM核心设施的能力 单粒子低温电子显微镜和低温电子断层扫描(CRYO-ET)中的SOM。目前，冷冻-EM的需求 大克利夫兰地区的研究社区由三台透射式电子显微镜支持： Tecnai T12、Tecnai TF20，以及最近添加的Krios G3i。通过Cryo-EM Core，用户可以从 CWRU、其地区性附属机构以及俄亥俄州东北部的其他大学。主要技术 低温EM的进步使这一方法更容易应用于不同的生物系统。 与NIH支持的冷冻-EM培训计划相结合，并增加对协作的机构支持 随着这些举措的实施，这些发展导致对低温EM的需求大幅增加。当我们的高中- 分辨率仪器是最先进的，我们的生产力受到老化筛查的严重影响 显微镜，尽管是根据持续服务合同进行维护的，但已越来越频繁和 修理期过长。拟议的冰川显微镜将满足七大 用户和八个次要用户。这台显微镜是我们目前200千伏显微镜的一大进步 具有更好的电子光学，柱子稳定性，更重要的是配备了更大容量的样品- 与Krios G3i直接兼容的搬运机器人。建议的工具的高级功能 对拟议实验的成功至关重要的因素包括：a)显著提高了对较小的- 样品大小对比度差；b)提高了Cryo-EM/Cryo-ET样品筛选、优化、 和高分辨率数据收集，以满足结构生物学家日益增长的需求；c)易于培训， 并更好地集成到现有的Cryo-EM工作流程中。美国国立卫生研究院资助的研究项目将直接 这种仪器的好处包括对一些临床相关靶点的结构研究，例如 离子通道、膜信号受体、普恩蛋白、免疫受体和病毒。总体而言， 收购新显微镜将扩大生物医学研究的范围，并进一步加强 我们在大克利夫兰地区的科学界内的多学科协作互动。

Project Summary/Abstract We propose to acquire a state-of-the-art 200 kV Glacios Cryo transmission electron microscope at Case Western Reserve University School of Medicine (CWRU SOM) for structural studies of biological macromolecular assemblies. Cryo-electron microscopy (Cryo-EM) has emerged as an unparalleled tool for high-resolution structure determination of macromolecules ranging from sizes as low as 40 kDa and as high as several MDa. The technology allows for imaging samples in solution, native membrane bilayers, or even in their physiological cellular environment. This new microscope will extend the capabilities of the Cryo-EM Core Facility at CWRU SOM in single-particle Cryo-EM and Cryo- electron tomography (Cryo-ET). Currently, the Cryo-EM needs of the research community in the Greater Cleveland area are supported by three transmission electron microscopes: Tecnai T12, Tecnai TF20, and most recently added Krios G3i. The Cryo-EM Core provides access to users from CWRU, its regional affiliate institutions, and other Universities in Northeast Ohio. The major technological advancements in Cryo-EM have made this methodology easier to adopt for diverse biological systems. Combined with NIH-supported Cryo-EM training programs and increased institutional support for collaborative initiatives, these developments have led to a major increase in the demand for pursing Cryo-EM. While our high- resolution instrumentation is state-of-the-art, our productivity is severely affected by our aging screening microscopes, which, although maintained under continued service contracts, have increasingly frequent and prolonged periods of repair. The proposed Glacios microscope will meet the substantial needs of seven major users and eight minor users. This microscope is a major advancement over our current 200 kV microscope featuring better electron optics, column stability, and more importantly is equipped with a larger capacity sample- handling robot that is directly compatible with the Krios G3i. Advanced features of the proposed instrument that are critical to the success of the proposed experiments include: a) significantly improved sensitivity for smaller- sized samples with poor contrast; b) enhanced throughput for Cryo-EM/Cryo-ET sample screening, optimization, and high-resolution data collection to cater to the growing needs of the structural biologists; c) ease of training, and better integration within the existing Cryo-EM workflow. The NIH-funded research projects that will directly benefit from this instrumentation include the structural studies of a number of clinically relevant targets such as ion channels, membrane signalling receptors, prion proteins, immune receptors, and viruses. Overall, the acquisition of the new microscope will expand the scope of biomedical research and further strengthen the multidisciplinary collaborative interactions within our scientific community here in the Greater Cleveland area.