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) 此应用程序寻求资金购买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，拥有NIH的活跃资金，以执行原子或近原子分辨率 粒子低温电子显微镜(CryoEM)研究。DDD摄像头将集成到现有的、多个 加州大学洛杉矶分校(UCLA)价值百万美元的泰坦·克里奥斯冷冻电子显微镜。这个 迫切需要一种能够记录大范围范围的原子分辨率低温EM图像的数字相机 具有生物医学意义的大分子复合体在多个层面上是合理的。这款DDD摄像头将提供 为13个需要原子级或近原子级结构测定的用户提供基本仪器 用低温电子显微镜进行分辨。目前，高分辨率的低温电磁图像是用过时的技术记录的： 图像记录在即将无法获得的照相胶片上；胶片必须扫描，需要 已经停产的许多工时和扫描仪。此外，胶片加载还引入了 进入显微镜柱的湿气，这会污染低温EM样品并减少显微镜的使用 时间(由于从柱中抽出)为~2倍。特别值得注意的是，因为用户不再 必须出现在显微镜前加载和卸载胶片，他们可以远程控制显微镜和 执行高分辨率的低温电磁成像，从而显著提高了对这款高端低温电磁成像设备的易用性 为南加州非加州大学洛杉矶分校的用户提供了一种工具，并降低了用户的成本。事实上，用户 可以通过夜间服务发送CryoEM网格，并且在我们的员工加载网格后，可以将他们的 远程采样长达3周。这13个用户来自南加州的领先机构，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)。所有这些用户都是由美国国立卫生研究院资助进行基础生物医学研究的， 生物化学、病毒学和微生物学。所有这些项目都使用了高分辨率的低温电磁成像技术。一起， 新的DDD相机将允许对来自这些生物样本的广泛范围的高分辨率低温EM 用户，使他们能够了解行动机制并确定新的发展目标 新疗法。此外，多样化的生物结构及其高度多样化的结构，提供了一种 为将高分辨率低温电子显微镜的极限提高到接近2？提供了丰富的数据来源。这些项目只是 代表一个潜在的更大的项目池，可以从进入这一高端市场中受益 乐器。因此，在加州大学洛杉矶分校的泰坦·克里奥斯仪器上安装DDD相机将大大 提高这些用户以及来自世界各地的潜在新用户的访问权限和效率，从而降低成本 世界。