Acquisition of a Focused Ion Beam Scanning Electron Microscope with cryo-stage

配备冷冻台的聚焦离子束扫描电子显微镜

• 批准号: 10415675
• 负责人: MATTHEW J TYSKA
• 金额: $ 163.86万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-15 至 2023-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10415675
• 关键词: 3-Dimensional; Academic Medical Centers; Architecture; Bacterial Proteins; Basement membrane; Biological; Businesses; Cancer Biology; Cell Volumes; Cell surface; Cells; Cellular Membrane; Collaborations; Communities; Complex; Congenital Abnormality; Cryo-electron tomography; Cryoelectron Microscopy; Development; Digestive System Disorders; Disease; Electron Microscope; Epithelial; Freezing; Funding; Health; Human; In Situ; Infrastructure; Intestinal Cancer; Investigation; Ions; Macromolecular Complexes; Maintenance; Malignant Neoplasms; Microscope; Modeling; Molecular Structure; Organelles; Performance; Protein Secretion; Request for Proposals; Research; Research Personnel; Research Project Grants; Resolution; Resource Sharing; Sampling; Scanning; System; Techniques; Thinness; Training; United States National Institutes of Health; Universities; cell assembly; cellular imaging; imaging capabilities; instrument; operation; reconstruction; structural biology; tomography; trafficking

Project Summary This proposal requests funds to acquire a Focused Ion Beam Scanning Electron Microscope (FIB-SEM) to support a diverse group of NIH-funded research projects at Vanderbilt University and Vanderbilt University Medical Center. This microscope will permit three-dimensional reconstruction of entire cell volumes with sufficient spatial resolution to observe the fine details of organelles. This volume information is essential to understand and quantify the complex interactions of organelles and macromolecular complexes within cells. The addition of a cryo-stage will extend the functionality of the FIB-SEM, adding ion milling of vitrified frozen cells to permit cryo-electron tomography of macromolecular structures in situ and by cryo-SEM. The Zeiss Crossbeam 550 is a state-of-the-art FIB-SEM with a high-performance ion column (3 nm resolution and up to 100 nA of probe current) that can efficiently perform FIB-SEM tomography and can be integrated with a Leica cryo-stage to create thin lamella for cryo-electron tomography. Our major users will apply FIB-SEM tomography to break new ground on NIH-funded research projects which span a wide range of problems in cell, developmental, epithelial, intestinal, and cancer biology, among others. Projects include studies of cytoskeletal dynamics (Tyska), assembly of cell surface features (Goldenring, Tyska), bacterial protein secretion (Lacy, Cover), basement membrane architecture (Weaver, Hudson), and cellular membrane trafficking (Weaver, Goldenring, Zhou). Together these investigations hold significance for our understanding of disease and illness related to birth defects, cancer, digestive disorders, and many other human health problems. The Crossbeam 550 will be a collaboration between the Vanderbilt Cell Imaging Shared Resource (CISR) and the Vanderbilt Cryo-EM Facility (V-CEM) within the Center for Structural Biology (CSB). It will be administered by the Vanderbilt Cell Imaging Shared Resource. The CISR will provide the infrastructure for introducing researchers across Vanderbilt to these new techniques, train users in operation of the instrument, and apply an established business model to support the long-term maintenance of the system. In summary, acquisition of the Zeiss Crossbeam 550 FIB- SEM will allow for unprecedented volumetric imaging capabilities of biological samples at scales and spatial resolutions not currently available to the Vanderbilt community and will facilitate research on a diverse set of topics.

项目概要 该提案要求资金购买聚焦离子束扫描电子显微镜 (FIB-SEM) 支持范德比尔特大学和范德比尔特大学由 NIH 资助的多元化研究项目 医疗中心。该显微镜将允许对整个细胞体积进行三维重建 足够的空间分辨率来观察细胞器的细节。此卷信息对于 理解和量化细胞内细胞器和大分子复合物的复杂相互作用。这 添加冷冻台将扩展 FIB-SEM 的功能，增加玻璃化冷冻细胞的离子研磨 允许通过冷冻扫描电镜对大分子结构进行原位冷冻电子断层扫描。蔡司横梁 550 是一款最先进的 FIB-SEM，配有高性能离子柱（3 nm 分辨率和高达 100 nA 的电流） 探针电流），可以有效地执行 FIB-SEM 断层扫描，并可以与徕卡冷冻台集成 创建用于冷冻电子断层扫描的薄层。我们的主要用户将应用FIB-SEM断层扫描来打破 NIH 资助的研究项目的新领域，涵盖细胞、发育、 上皮、肠道和癌症生物学等。项目包括细胞骨架动力学研究 (Tyska)、细胞表面特征的组装(Goldenring、Tyska)、细菌蛋白分泌(Lacy、Cover)、 基底膜结构（Weaver、Hudson）和细胞膜运输（Weaver、Goldenring、 周）。这些研究对于我们了解与疾病相关的疾病具有重要意义 先天缺陷、癌症、消化系统疾病和许多其他人类健康问题。 Crossbeam 550 将是 范德比尔特细胞成像共享资源 (CISR) 和范德比尔特冷冻电镜之间的合作 结构生物学中心 (CSB) 内的设施 (V-CEM)。它将由范德比尔特小组管理 成像共享资源。 CISR 将为在范德比尔特引入研究人员提供基础设施 针对这些新技术，培训用户操作仪器，并将已建立的商业模式应用于 支持系统的长期维护。总之，购买 Zeiss Crossbeam 550 FIB- SEM 将实现生物样品在尺度和空间上前所未有的体积成像能力 范德比尔特社区目前尚未获得决议，并将促进对各种不同问题的研究 主题。