ICP-QQQ

项目摘要 最近的方法和技术进步导致了电子显微镜的迅速发展。 (Em)加州大学伯克利分校和更广泛的科学界的应用。特别是，单粒子冷冻-EM 它的应用在结构生物学领域带来了许多引人注目的进展。同样引人注目的是 低温电子层析成像(CRYO-ET)和体积电磁场的平行场，两者都是由于聚焦离子的进步而实现的 光束(FIB)铣削，在复杂生物系统的研究中已日益成为主流。这些 方法允许直接高分辨率可视化的大分子结构和细胞结构在一个本机 在此背景下，以前所未有的方式将细胞生物学和结构生物学领域联系起来。加州大学伯克利分校 电子显微镜和细胞生物学领域长期以来一直处于各自领域的前沿，现在已经开始 基于对结合这些有前景的原位EM可视化方法的持续投资。作为研究 植根于这些方法的项目已经开始成熟，需要我们的教员获得尖端技术 仪表化已经变得既紧迫又非常明确。目前，还没有高端的冷冻机 光纤扫描电子显微镜(CRYO-FIB-SEM)可供我校科研人员使用， 生物样本。在这份提案中，我们申请资金购买一架低温蔡司横梁(XB)550 纤维扫描电子显微镜和辅助低温仪器。这台仪器的尖端性能将大大提升 加州大学伯克利分校致力于刻画生物结构的多个数量级，从埃到埃 毫米长的天平。蔡司XB 550FIB-SEM显微镜能够确定3D体积-EM 在房间(3 nm分辨率，树脂包埋)和低温(30 nm分辨率)下成像的细胞结构 温度。凭借其在低温FIB铣削方面的尖端能力，蔡司XB 550也将无缝 集成到校园低温管道中，为以后的高分辨率低温发射产生细胞低温薄片 使用伯克利现有的两台泰坦-克里奥斯冷冻电子显微镜进行成像。购置拟议的系统 将极大地影响三大用户和五小用户群体的工作，这三大用户群体和五个小用户群体专注于重要的 关于病毒复制和发病机制、细胞信号和营养的生物和生物医学问题 摄取，肿瘤发生，纤毛功能障碍，细胞器生物发生。加州大学伯克利分校的电子显微镜 实验室，由经验丰富的生物显微镜专家组成，他们曾为校园学术人员服务 研究人员几十年来，将安装蔡司XB 550。我们收购蔡司XB 550将完全 使我们的新兴市场基础设施现代化，加快加州大学伯克利分校在可持续发展道路上的进展 细胞和分子生物学的根本性突破。

Project Summary Recent methodological and technological advances have resulted in a rapid expansion of electron microscopy (EM) applications at UC Berkeley and in the wider scientific community. In particular, single particle cryo-EM applications have led to numerous high-profile advances in the field of structural biology. The equally compelling parallel fields of cryo-electron tomography (cryo-ET) and volume EM, both enabled by advances in focused ion beam (FIB) milling, have become increasingly mainstream in the study of complex biological systems. These methods allow for the direct high-resolution visualization of macromolecular and cellular structures within a native context, bridging the fields of cell biology and structural biology in an unprecedented manner. The UC Berkeley electron microscopy and cell biology communities, long at the forefront of their respective fields, have embarked upon a sustained investment in combining these promising in situ EM visualization methods. As research projects grounded in these methods have begun to mature, the need for our faculty to access cutting-edge instrumentation has become both urgent and abundantly clear. Currently, there exists no high-end cryo-capable FIB scanning electron microscope (cryo-FIB-SEM) accessible to researchers on our campus with complex, biological samples. In this proposal, we request funds to purchase a cryo-capable Zeiss Crossbeam (XB) 550 FIB-SEM and ancillary cryo-instrumentation. The cutting-edge capabilities of this instrument will greatly advance efforts at UC Berkeley to characterize biological structures across many orders of magnitude, from angstrom to millimeter length scales. The Zeiss XB 550 FIB-SEM microscope enables the determination of 3D volume-EM imaged cellular structures at both room (3 nm resolution, resin-embedded) and cryo (30 nm resolution) temperatures. With its cutting-edge capabilities in cryo-FIB milling, the Zeiss XB 550 will also be seamlessly integrated into the campus cryo-pipeline to generate cellular cryo-lamellae for later high resolution cryo-ET imaging using Berkeley’s two existing Titan Krios cryo-electron microscopes. Acquisition of the proposed system will greatly impact the work of three Major User and five Minor User groups, which are focused on important biological and biomedical questions regarding viral replication and pathogenesis, cellular signaling and nutrient uptake, oncogenesis, cilia dysfunction, and organelle biogenesis. The UC Berkeley Electron Microscopy Laboratory, expertly staffed by experienced biological microscopists who have served campus academic researchers for decades, will house the Zeiss XB 550. Our acquisition of the Zeiss XB 550 will completely modernize our EM infrastructure, accelerating UC Berkeley’s progress on a sustainable path to drive fundamental breakthroughs in cell and molecular biology.