Acquisition of 200kV Glacios Cryo Transmission Electron Microscope

购置 200kV Glacios 冷冻透射电子显微镜

• 批准号: 10430469
• 负责人: Sudha Chakrapani
• 金额: $ 200万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-15 至 2024-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10430469
• 关键词: Adopted; Affect; Aging; Area; Biological; Biomedical Research; Communities; Contract Services; Core Facility; Cryo-electron tomography; Cryoelectron Microscopy; Data Collection; Development; Electron Microscope; Environment; Felis catus; Funding; Image; Immunologic Receptors; Institution; Ion Channel; Membrane; Methodology; Microscope; Minor; Ohio; Physiological; PrP; Productivity; Receptor Signaling; Research; Research Project Grants; Resolution; Robot; Sample Size; Sampling; Structural Biologist; Structure; Technology; Training; Training Programs; United States National Institutes of Health; Universities; Virus; biological systems; clinically relevant; electron optics; experimental study; improved; instrument; instrumentation; macromolecular assembly; macromolecule; medical schools; multidisciplinary; particle; repaired; screening; success; tool; transmission process; Adopted; Affect; Aging; Area; Biological; Biomedical Research; Communities; Contract Services; Core Facility; Cryo-electron tomography; Cryoelectron Microscopy; Data Collection; Development; Electron Microscope; Environment; Felis catus; Funding; Image; Immunologic Receptors; Institution; Ion Channel; Membrane; Methodology; Microscope; Minor; Ohio; Physiological; PrP; Productivity; Receptor Signaling; Research; Research Project Grants; Resolution; Robot; Sample Size; Sampling; Structural Biologist; Structure; Technology; Training; Training Programs; United States National Institutes of Health; Universities; Virus; biological systems; clinically relevant; electron optics; experimental study; improved; instrument; instrumentation; macromolecular assembly; macromolecule; medical schools; multidisciplinary; particle; repaired; screening; success; tool; transmission process

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.

项目摘要/摘要 我们建议在Case Western处获得最先进的200 kV冰川冷冻传输电子显微镜 预备大学医学院（CWRU SOM）用于生物大分子的结构研究 集会。冷冻电子显微镜（Cryo-EM）已成为高分辨率的无与伦比的工具 大分子的结构测定范围从低至40 kDa和高达几个MDA的大小。 该技术允许在溶液，本地膜双层甚至其生理学中成像样品 细胞环境。这种新的显微镜将扩展CWRU中的Cryo-EM核心设施的功能 SOM在单粒子冷冻EM和冷冻电子断层扫描（Cryo-ET）中。目前，Cryo-Em的需求 大克利夫兰地区的研究社区得到了三个传输电子显微镜的支持： Tecnai T12，Tecnai TF20，最近添加了Krios G3i。 Cryo-Em Core提供了从 CWRU，其区域会员机构和俄亥俄州东北部的其他大学。主要技术 Cryo-EM的进步使该方法更容易用于多种生物系统。 结合NIH支持的冷冻EM培训计划，并增加了对协作的机构支持 这些事态发展导致追求冷冻EM的需求大大增加。而我们的高 解决方案仪器是最先进的，我们的生产力受到老化筛查的严重影响 显微镜虽然在持续的服务合同下维持，但越来越频繁，并且 长时间的维修期。拟议的冰川显微镜将满足七个主要的大量需求 用户和八个次要用户。该显微镜是我们当前200 kV显微镜的重大进步 具有更好的电子光学元件，柱稳定性，更重要的是配备更大的容量样品 - 处理与Krios G3i直接兼容的机器人。提议的乐器的高级功能 对于拟议的实验的成功至关重要，包括：a）显着提高较小的灵敏度 大小的样品，对比度不佳； b）增强了Cryo-EM/Cryo-ET样品筛选，优化，增强的吞吐量 和高分辨率数据收集，以满足结构生物学家不断增长的需求； c）易于培训， 并更好地集成在现有的Cryo-EM工作流程中。 NIH资助的研究项目将直接 该仪器的好处包括对许多临床相关目标的结构研究，例如 离子通道，膜信号受体，prion蛋白，免疫受体和病毒。总体而言， 新显微镜的获取将扩大生物医学研究的范围，并进一步加强 大克利夫兰地区的科学界内部的多学科合作互动。