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） 该应用程序寻求资金购买67百万像素直接检测设备（DDD）相机以支持 南加州的13位用户拥有活跃的NIH资金，以执行原子或近原子分辨率单。 颗粒冷冻电子显微镜（冷冻）研究。 DDD摄像机将集成到现有的多个 洛杉矶分校（UCLA）的百万美元泰坦KRIOS CRYO ELECTRON显微镜。 对能够录制原子分辨率冷冻图像的数码相机的批判性需求 生物医学意义的大分子复合物在多个水平上是合理的。这款DDD相机将提供 对于需要在原子或靠近原子的13位用户的必需仪器 冷冻解决方案。目前，使用过时的技术记录高分辨率冷冻图像： 图像记录在摄影电影中，这些电影将很快不可用；电影必须扫描，需要 许多已经停产的人小时和扫描仪。此外，胶片负载介绍 水分进入显微镜柱，该柱污染冷冻样品并降低可用显微镜 时间（由于从列中抽出）〜2。特别要注意，因为用户将不再 必须在显微镜上存在加载和卸载膜，它们可以远程控制显微镜和 执行高分辨率的冷冻成像，从而显着提高了该高端冷冻的可访问性 南加州的非美国联社用户的仪器，并降低了用户的成本。确实，用户 可以通过隔夜服务发送冷冻电网，并且在我们的员工加载网格后，可以对其进行想象 远程样品最多3周。 13位用户来自南加州的领先机构，包括 南加州大学加州大学加州大学加州大学加州大学科技研究所（USC）和Scripps研究 研究所（Scripps）。所有这些用户都是由NIH资助的，用于购买基本生物医学研究的研究， 生物化学，病毒学和微生物学。所有项目都采用高分辨率冷冻成像。一起， 新的DDD摄像头将允许使用这些摄像机的大量生物样品允许高分辨率冷冻 用户，使他们能够理解行动机制，并确定开发的新目标 新疗法。此外，潜水员生物结构及其高度多样的建筑提供了一种 将高分辨率冷冻限制到接近2的数据来源。这些项目只是 代表可能受益于进入此高端的可能受益的更大的项目池 乐器。因此，在UCLA的Titan Krios仪器中安装DDD摄像头会极大地 提高访问，效率，从而降低这些用户的成本，并可能来自各地的新用户 世界。