West/Midwest Consortium for High-Resolution Cryo Electron Microscopy

标题：西部/中西部高分辨率低温电子显微镜联盟 总结 该申请寻求资金，以建立西部/中西部高分辨率低温电子联盟 显微镜检查（cryoEM）。该联盟的目标是免费为来自10个地区研究所的19名cryoEM用户提供服务。 进入位于加州的高端cryoEM设施， 加州大学洛杉矶分校（UCLA）的纳米系统研究所（CNSI）。加州大学洛杉矶分校将作为主办机构 并为这些cryoEM实验室的研究人员提供其高产的Titan Krios cryo electron 最近升级的显微镜，配有Volta相位板，Gatan成像滤波器（GIF），以及GIF前后 直接电子探测器一个经验丰富的工作人员与证明记录的cryoEM衍生原子结构 将提供现场协助，以收集数据，并提供精简的“电影”预处理，为我们的 联盟用户实验室。该联盟记录原子分辨率cryoEM的关键需求 广泛的生物医学意义上的大分子复合物的图像是合理的， 为19个用户提供服务。特别值得注意的是，我们已经证明，一旦经过我们工作人员的培训， 用户不再需要在显微镜前，可以远程控制显微镜并执行 在我们的工作人员将仪器对准以获得最佳性能后， 大大提高了高端仪器的可访问性。三人监督委员会 代表非联合体用户、区域联合体用户和UCLA设施工作人员将提供 评估和批准用户项目，添加和删除用户。在我们的典型操作下，用户组 与批准的项目研究仪器时间与现有的专业设计的在线预订 系统，通过隔夜快递（例如FedEx）在液氮冷却的干燥运输箱中发送cryoEM网格，以及 在我们的工作人员加载他们的网格后，远程成像他们的样品平均3-7天，最多2-3天 如果有必要的话，几周。这10个研究所集中在西海岸（西雅图的华盛顿大学， 加州大学圣地亚哥分校，斯克里普斯）和中西部（凯斯西储大学，普渡 大学、堪萨斯大学、伯明翰的亚拉巴马大学），但三所大学除外（ 得克萨斯州埃尔帕索，得克萨斯大学医学分支在加尔维斯顿，和佛罗里达大学盖恩斯维尔）。 除了三个新的研究者，所有这些用户都是由NIH或其他联邦机构资助的 从事生物医学研究，包括生物学、生物化学、病毒学和微生物学。的 该联盟的成立将立即赋予我们的用户高分辨率cryoEM能力 为广泛的生物样品，使他们能够了解作用机制，并确定新的 用于开发新疗法的靶点。

Title: West/Midwest Consortium for High-Resolution Cryo Electron Microscopy Summary This application seeks funding to establish the West/Midwest Consortium for High-Resolution Cryo Electron Microscopy (cryoEM). The goal of the consortium is to offer 19 cryoEM users from 10 regional institutes free access to a high-end cryoEM facility with proven high-resolution capabilities located in the California NanoSystems Institute (CNSI) at University of California, Los Angeles (UCLA). UCLA will act as host institute and provide investigators in these cryoEM laboratories access to its highly productive Titan Krios cryo electron microscope recently upgraded with a Volta phase plate, a Gatan imaging filter (GIF), and pre- and post-GIF direct electron detectors. A highly experienced staff with proven records of cryoEM-derived atomic structures will provide on-site assistance to collect data and provide streamlined “movie” pre-processing for our consortium user laboratories. The critical need for this consortium for recording atomic-resolution cryoEM images of a broad range of biomedically significant macromolecular complexes is justified by the immediate benefits to the 19 consortium users. Of particular note, we have demonstrated that, once trained by our staff, users no longer have to be present at the microscope and can remotely control the microscope and perform high-resolution cryoEM imaging after our staff have aligned the instruments for top performance, thus significantly improving accessibility to the high-end instrument. A three-member supervisory committee representing non-consortium users, regional consortium users and the UCLA facility staff will provide evaluation and approval of user projects, adding and dropping users. Under our typical operation, a user group with approved projects research instrument time with an existing professionally-designed online reservation system, sends cryoEM grids by overnight courier (such as FedEx) in a liquid nitrogen-cooled dry shipper, and after their grids are loaded by our staff, image their sample remotely for 3-7 days on average, and up to 2-3 weeks if necessary. The 10 institutes are concentrated on the West coast (University of Washington in Seattle, University of California at San Diego, Scripps) and Midwest (Case Western Reserve University, Purdue University, The University of Kansas, University of Alabama at Birmingham) except for three (University of Texas at El Paso, University of Texas Medical Branch in Galveston, and University of Florida at Gainesville). With the exception of three new investigators, all these users are funded by the NIH or other federal agencies to pursue biomedical research, ranging from biology, biochemistry, virology and microbiology. The establishment of this consortium will immediately empower our users with high-resolution cryoEM capabilities for a broad range of biological samples, enabling them to understand mechanisms of action and identify new targets for the development of new therapeutics.