Administrative Supplement: The Midwest Center for Cryo-Electron Tomography

修改后的项目摘要/摘要部分 我们建议在威斯康星大学麦迪逊分校的生物化学系建立一个冷冻电子断层扫描的全国服务网络中心。中西部低温电子断层扫描中心(MCCET)将为调查人员提供以下方面的支持：(1)常规和高级低温电子断层扫描(CRYO-ET)样本准备、数据收集和计算；以及(2)低温电子断层扫描样本准备、数据收集、数据处理和验证方面的实际、远程和虚拟培训。MCCET的业务将建立在威斯康星大学麦迪逊分校生物化学系建立的冷冻-EM研究中心(CEMRC)的基础上。威斯康星大学麦迪逊分校在致力于电子显微镜以及电子显微镜在生物和材料科学方面的进步方面有着悠久的历史。安装在MCCET中的辅助设备和主要仪器将包括：细胞培养设备、荧光显微镜和微型图案化系统、辉光放电设备、碳蒸发器、自动插入式冷冻机、高压冷冻机、低温超微切割机、低温聚焦离子束扫描电子显微镜(CRYO-FIB-SEM)和一台300千伏的透射电子显微镜。联系人伊丽莎白·R·赖特博士拥有20多年的低温EM经验，包括开发新的低温EM技术和工作流程，以及指导和管理(低温)电子显微镜研究资源。Marisa Otegui博士、Robert Kirchdoerfer博士和Tim Grant博士带来了复杂组织、小分子和细胞器的样本保存和成像、成像和计算技术开发以及设施管理方面的互补专业知识。一名中心经理、一名计算机科学家/系统管理员、一名项目经理、一名低温技术培训讲师、一名计算机科学家和博士后研究助理将管理MCCET的日常操作，并支持用户完成其项目的整个过程，从初始访问、样品处理和准备、样品验证、低温技术数据收集和处理以及交叉培训。中心与NIH、网络中心和用户的协调活动将由PI、项目经理、中心经理和计算机科学家/系统管理员管理。中西部低温电子断层扫描中心(MCCET)将在低温电子断层扫描领域保持一种创新和技术开发的文化，以促进冷藏样品制备、产量、验证和成像方面的改进。MCCET将是一个以用户体验为重点的资源，将为许多学科的研究人员提供访问和培训。

Modified Project Summary/Abstract Section We propose to establish a National Service Network Hub for Cryo-Electron Tomography to be hosted in the Department of Biochemistry at the University of Wisconsin, Madison. The Midwest Center for Cryo-Electron Tomography (MCCET) will support investigators by providing access to well-trained staff and state-of-the-art equipment for (1) routine and advanced cryo-electron tomography (cryo-ET) specimen preparation, data collection, and computation; and (2) hands-on, remote, and virtual training in cryo-ET specimen preparation, data collection, and data processing and validation. MCCET operations will build on the Cryo-EM Research Center (CEMRC) established in the Department of Biochemistry at the University of Wisconsin, Madison. The University of Wisconsin, Madison has a long history of institutional commitment to electron microscopy, as well as the advancement of electron microscopy in both the biological and materials sciences. The ancillary equipment and major instrumentation to be housed in the MCCET will include: cell culture equipment, a fluorescence light microscope and micropatterning system, glow discharge devices, carbon evaporator, automated plunge freezer, high pressure freezer, cryo-ultramicrotome, a cryo-focused ion beam-scanning electron microscope (cryo-FIB-SEM), and a 300 kV TEM. Dr. Elizabeth R. Wright, the contact PI, has over twenty-years of cryo-EM experience including the development of new cryo-EM technologies and workflows along with directing and managing (cryo-)electron microscopy research resources. Drs. Marisa Otegui, Robert Kirchdoerfer, and Tim Grant bring complementary expertise in sample preservation and imaging of complex tissues, small molecules and organelles, imaging and computational technology development, as well as facility management. A center manager, a computer scientist/systems administrator, a project manager, cryo- ET training instructors, a computer scientist, and postdoctoral research associates will manage MCCET day-today operations and support users through the course of their projects from initial access, sample handling and preparation, sample validation, cryo-ET data collection and processing, and cross-training. Hub coordination activities with the NIH, network centers, and users will be managed by the PI, project manager, center manager, and computer scientist/systems administrator. The Midwest Center for Cryo-Electron Tomography (MCCET) will maintain a culture of innovation and technology development in cryo-ET to facilitate improvements to cryo-ET sample preparation, throughput, validation, and imaging. The MCCET will be a user-experience focused resource that will provide access and training to researchers across many disciplines.