The Midwest Center for Cryo-Electron Tomography

修改项目摘要/摘要部分 我们建议建立一个国家服务网络中心的低温电子断层扫描在生物化学系在威斯康星州，麦迪逊的大学。中西部冷冻电子断层扫描中心（MCCET）将通过提供训练有素的工作人员和最先进的设备来支持研究者，这些设备用于（1）常规和高级冷冻电子断层扫描（cryo-ET）标本制备、数据收集和计算;以及（2）在冷冻ET标本制备、数据收集以及数据处理和验证方面的动手、远程和虚拟培训。MCCET的运作将建立在麦迪逊的威斯康星州大学生物化学系建立的冷冻EM研究中心（CEMRC）的基础上。威斯康星州，麦迪逊的大学有着悠久的历史，致力于电子显微镜的机构，以及电子显微镜在生物和材料科学的进步。MCCET中的辅助设备和主要仪器包括：细胞培养设备、荧光显微镜和微图案系统、辉光放电装置、碳蒸发器、自动插入式冷冻机、高压冷冻机、冷冻超薄切片机、冷冻聚焦离子束扫描电子显微镜（cryo-FIB-SEM）和300 kV TEM。伊丽莎白·R博士Wright是联系人PI，拥有超过20年的cryo-EM经验，包括开发新的cryo-EM技术和工作流程沿着指导和管理（cryo-）电子显微镜研究资源。Marisa Otegui、Robert Kirchdoerfer和Tim Grant博士在复杂组织、小分子和细胞器的样品保存和成像、成像和计算技术开发以及设施管理方面具有互补的专业知识。中心经理、计算机科学家/系统管理员、项目经理、cryo-ET培训讲师、计算机科学家和博士后研究助理将管理MCCET日常操作，并在项目过程中为用户提供支持，包括初始访问、样品处理和制备、样品验证、cryo-ET数据收集和处理以及交叉培训。与NIH、网络中心和用户的中心协调活动将由PI、项目经理、中心经理和计算机科学家/系统管理员管理。中西部冷冻电子断层扫描中心（MCCET）将保持冷冻电子断层扫描的创新和技术开发文化，以促进冷冻电子断层扫描样品制备、通量、验证和成像的改进。MCCET将是一个以用户体验为中心的资源，将为多个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.