HIGH-END CRYOEM INSTRUMENT FOR THE UCLA ELECTRON IMAGING CENTER FOR NANOMACHINES

该子项目是利用由NIH/NCRR资助的共享仪器补助金提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是资助机构，不一定是研究者的机构。加州大学洛杉矶分校（UCLA）的加州纳米系统研究所（CNSI）的长期目标是为尖端的纳米生物学和生物医学研究创造一个有利的跨学科环境。作为这一目标的一个组成部分，纳米机器电子成像中心（EICN）的建立是为了满足在纳米尺度上可视化大分子机械和理解其作用机制的先进成像技术的迫切需要。这项高端共享仪器提案寻求50%的资金（160万美元），用于为EICN购买一台SOOkV场发射枪电子显微镜，以增强我们正在进行的三维（3D）结构研究，通过电子冷冻显微镜（cryoEM）和断层扫描（ET）对广泛的纳米机械、细胞器、病毒和细菌细胞进行研究。对用户友好、高分辨率、具有层析成像能力的电子冷冻显微镜仪器的迫切需求在多个层面上是合理的：高分辨率电子显微镜将为nih资助的研究项目的十几个主要用户提供必要的仪器，这些项目需要通过低温电子显微镜在亚纳米分辨率或低温电子显微镜在分子分辨率下进行结构测定。目前，加州大学洛杉矶分校校园内没有工作的低温低温仪器。加州理工学院附近帕萨迪纳地区的低温电子冷冻显微镜由于当地用户已经满负荷运行，加州大学洛杉矶分校的教职员工无法使用。因此，通过获得应用中描述的高端电子低温显微镜，ucla的研究将大大加强。高分辨率电子成像将成为加州大学洛杉矶分校已经建立的非常强大的结构生物学研究社区的一个组成部分。新的高端EM仪器，加上完善的x射线晶体学，高分辨率核磁共振光谱学和分子建模专业知识，将为渴望扩大其当前生物医学和纳米生物学研究项目范围的教师提供宝贵的资源，包括低温EM和低温et。新仪器还将满足高分辨率数据的关键要求，以推动低温电镜重建的极限达到近原子分辨率。不同的生物结构及其高度多样化的结构为开发和完善高分辨率低温电子显微镜的方法提供了丰富的数据来源，这不仅有利于我们的努力，也有利于整个电子成像界。联邦政府积极资助的次要/次要用户的身份表明，自然科学学院和工程学院以及加州大学洛杉矶分校医学院的多个部门/研究所都对这种高端仪器有很大兴趣，它们可能会从购买这种高端仪器中受益匪浅。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): The long-term goal of the California NanoSystems Institute (CNSI) at the University of California at Los Angeles (UCLA) is to create an enabling and cross-disciplinary environment for cutting-edge nanobiology and biomedical research. As an integral part of this goal, the Electron Imaging Center for Nanomachines (EICN) is established to meet the pressing need of advanced imaging techniques for visualizing macromolecular machineries at the nanometer scale and for the understanding of their mechanisms of action. This high-end shared instrumentation proposal seeks 50% of the funds ($1.6M) needed for the purchase of a SOOkV field emission gun electron microscope for EICN to augment our on-going three- dimensional (3D) structural studies of a broad range of nanomachineries, organelles, viruses and bacterial cells by electron cryomicroscopy (cryoEM) and tomography (ET). The critical need for a user-friendly, high- resolution, tomography-capable electron cryomicroscopy instrument is justified at multiple levels: The high-resolution electron microscope will provide essential instrumentation for over a dozen major users with NIH-funded research projects that require structure determinations either at subnanometer resolution by cryoEM or at molecular resolution by cryoET. Currently, there is no working cryoEM instrument on the UCLA campus. The cryoET-capable electron cryomicroscope in the neighboring Pasadena area at Caltech is not accessible to UCLA faculty members due to already full-capacity operation by local users there. Therefore, UCLA-based research would be greatly enhanced by the acquisition of the high-end electron cryomicroscope described in the application. High-resolution electron imaging will become an integral part of the very strong structural biology research community already established at UCLA. The new high-end EM instrumentation, together with the well-established X-ray crystallography, high-resolution NMR spectrometry, and the molecular modeling expertise will provide a valuable resource for faculty members who are eager to expand the scope of their current biomedical and nanobiology research projects to include cryoEM and cryoET. The new instrument will also meet a critical requirement of high-resolution data for pushing the envelope of cryoEM reconstruction to near atomic resolution. The diverse biological structures with their highly varied architectures offer a fertile source of data for developing and refining the methodology of high-resolution cryoEM, which will not only benefit our efforts but also the general electron imaging community at large. The identification often secondary/minor users with active federal funding shows that there is major interest across multiple departments/institutes among the colleges of natural sciences and engineering, as well as the UCLA medical school that may greatly benefit from the acquisition of the high-end instrument.