West/Midwest Consortium for High-Resolution Cryo Electron Microscopy

 描述(由申请人提供)：本申请寻求资金，以建立西部/中西部高分辨率低温电子显微镜(CryoEM)联盟。该联盟的目标是让来自10个地区研究所的19名低温EM用户免费使用位于加州大学洛杉矶分校(UCLA)加州纳米系统研究所(CNSI)的高端低温EM设施，该设施具有公认的高分辨率能力。加州大学洛杉矶分校将作为主办机构，为这些低温EM实验室的研究人员提供其高产的泰坦Krios冷冻电子显微镜，最近升级了Volta相位板、Gatan成像滤光片(GIF)以及GIF前后的直接电子探测器。经验丰富的工作人员将提供现场协助，收集数据，并为我们联盟的用户实验室提供简化的“电影”前处理。该联盟迫切需要记录一系列具有生物医学意义的大分子复合体的原子分辨率低温电子显微镜图像，19个联盟用户的直接利益证明了这一点。特别值得注意的是，我们已经证明，一旦我们的员工进行了培训，用户就不必再出现在显微镜前，可以远程控制显微镜，并在我们的员工调整仪器以实现最佳性能后执行高分辨率的低温电磁成像，从而显著改善了高端仪器的可及性。一个由三人组成的监督委员会将代表非财团用户、地区财团用户和加州大学洛杉矶分校设施工作人员，对用户项目进行评估和批准，增加和删除用户。在我们的典型操作下，拥有批准项目的用户组要求使用现有的专业设计的在线预订系统来获得研究仪器的时间，通过隔夜快递(如联邦快递)在液氮冷却的干燥托运人中发送CryoEM网格，然后，在我们的工作人员加载网格后，对其样本进行远程成像，平均3-7天，如果需要，最长可达2-3周。这10个研究所集中在西海岸(西雅图华盛顿大学、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、阿拉巴马大学伯明翰分校)，除了三个(德克萨斯大学埃尔帕索分校、加尔维斯顿德克萨斯大学医学分校和佛罗里达大学盖恩斯维尔分校)。除了三名新的研究人员外，所有这些用户都是由美国国立卫生研究院或其他联邦机构资助进行生物医学研究，范围从生物学、生物化学、病毒学和微生物学。该联盟的成立将立即增强我们的用户对各种生物样本的高分辨率冷冻EM能力，使他们能够了解作用机制并确定新疗法开发的新靶点。

 DESCRIPTION (provided by applicant): 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 bio-medically 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 requests 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.