Direct Detection Device for atomic resolution cryoEM of macromolecular complexes

大分子复合物原子分辨率冷冻电镜直接检测装置

• 批准号: 8640787
• 负责人: Z Hong ZHOU
• 金额: $ 59.79万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-20 至 2015-04-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8640787
• 关键词: Architecture; Biochemistry; Biological; Biomedical Research; California; Cryoelectron Microscopy; Data Sources; Detection; Development; Devices; Electron Microscope; Film; Funding; Hour; Image; Imaging technology; Institutes; Los Angeles; Macromolecular Complexes; Microbiology; Microscope; Persons; Production; Pump; Research; Research Institute; Resolution; Sampling; Scanning; Services; Structure; Technology; Time; Titan; United States National Institutes of Health; Universities; base; cost; digital; improved; instrument; instrumentation; novel therapeutics; particle; virology

Project Summary (line 7) This application seeks funding to purchase a 67 megapixel Direct Detection Device (DDD) camera to support 13 users in Southern California with active NIH funding to carry out atomic or near-atomic resolution single particle cryo electron microscopy (cryoEM) studies. The DDD camera will be integrated into an existing, multi- million-dollar Titan Krios cryo electron microscope at the University of California, Los Angeles (UCLA). The critical need for a digital camera capable of recording atomic-resolution cryoEM images of a broad range of biomedically significant macromolecular complexes is justified at multiple levels. This DDD camera will provide essential instrumentation for 13 users that require structure determinations at either atomic or near atomic resolution by cryoEM. Currently, high resolution cryoEM images are recorded with obsolete technology: images are recorded on photographic film that will soon to be unavailable; the films must be scanned, requiring many person-hours and scanners that are already out of production. In addition, film-loading introduces moisture into the microscope column, which contaminates cryoEM samples and reduces usable microscope time (due to pumping out of the column) by a factor of ~2. Of particular note, because users would no longer have to be present at the microscope to load and unload film, they can remotely control the microscope and perform high-resolution cryoEM imaging, thus significantly improving accessibility to this high-end cryoEM instrument for non-UCLA-based users in Southern California, and reducing cost to the users. Indeed, users could send cryoEM grids by overnight service, and, after their grids are loaded by our staff, could image their sample remotely for up to 3 weeks. The 13 users are from leading institutes in Southern California, including UCLA, California Institute of Technology, University of Southern California (USC) and the Scripps Research Institute (Scripps). All of these users are funded by the NIH to pursue research in basic biomedical research, biochemistry, virology and microbiology. All of the projects employ high-resolution cryoEM imaging. Together, the new DDD camera will allow high-resolution cryoEM for a broad range of biological samples from these users, enabling them to understand mechanisms of actions and to identify new targets for the development of new therapeutics. Moreover, the diverse biological structures, with their highly varied architectures, offer a fertile source of data for pushing the limit of high-resolution cryoEM to near 2¿. These projects are only representative of a potentially much larger pool of projects that could benefit from access to this high-end instrument. Therefore, the installation of the DDD camera in the Titan Krios instrument at UCLA would greatly enhance the access, efficiency and thus reduce cost of these users and potentially new users from all over the world.

项目摘要（第7行） 这项申请寻求资金购买6700万像素的直接检测设备（DDD）摄像头，以支持 南加州的13个用户，NIH积极资助，进行原子或近原子分辨率的单 粒子冷冻电子显微镜（cryoEM）研究。DDD摄像头将集成到现有的多功能 在加州大学洛杉矶（UCLA），价值百万美元的泰坦克里奥低温电子显微镜。的 迫切需要一种能够记录宽范围的原子分辨率cryoEM图像的数码相机， 生物医学上重要的大分子复合物在多个水平上是合理的。这台DDD相机将提供 为13个需要在原子级或近原子级进行结构测定的用户提供必要的仪器 cryoEM分辨率。目前，使用过时的技术记录高分辨率cryoEM图像： 图像被记录在不久将无法获得的摄影胶片上;胶片必须被扫描， 许多人工小时和扫描仪已经停产。此外，胶片加载还引入了 水分进入显微镜柱，污染cryoEM样本，减少可用的显微镜 时间（由于从色谱柱中泵出）缩短约2倍。特别值得注意的是，由于用户不再 必须在场的显微镜加载和卸载电影，他们可以远程控制显微镜， 执行高分辨率cryoEM成像，从而显著提高这种高端cryoEM的可访问性 仪器为非加州大学洛杉矶分校的用户在南加州，并降低成本的用户。事实上，用户 可以通过夜间服务发送cryoEM网格，并且，在我们的工作人员加载网格后，可以对他们的网格进行成像。 远程采样长达3周。这13名用户来自南加州的领先研究所，包括 加州大学洛杉矶分校、加州理工学院、南加州大学（USC）和斯克里普斯研究所 研究所（斯克里普斯）。所有这些用户都由NIH资助，从事基础生物医学研究， 生物化学、病毒学和微生物学。所有项目都采用高分辨率cryoEM成像。在一起， 新的DDD相机将允许高分辨率的冷冻EM用于广泛的生物样品， 用户，使他们能够了解行动机制，并确定新的目标， 新疗法此外，生物结构的多样性及其高度变化的结构， 将高分辨率cryoEM的极限推到近2 º的丰富数据来源。这些项目只是 代表了一个潜在的更大的项目池，可以受益于获得这一高端 仪器因此，在加州大学洛杉矶分校的Titan Krios仪器中安装DDD相机将大大提高 提高这些用户和来自世界各地的潜在新用户的访问、效率，从而降低成本。 世界