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.

项目摘要 该提案要求提供资金，以购置一台聚焦离子束扫描电子显微镜， 支持范德比尔特大学和范德比尔特大学的各种NIH资助的研究项目 医学中心这种显微镜将允许整个细胞体积的三维重建， 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。的 冷冻台的增加将扩展FIB-SEM的功能，增加玻璃化冷冻细胞的离子研磨， 允许原位和通过cryo-SEM对大分子结构进行低温电子断层扫描。蔡司Crossbeam 550是最先进的FIB-SEM，具有高性能离子柱（3 nm分辨率和高达100 nA的 探头电流），可有效执行FIB-SEM断层扫描，并可与Leica冷冻台集成 为低温电子断层扫描制作薄层。我们的主要用户将应用FIB-SEM断层扫描来打破 美国国立卫生研究院资助的研究项目的新领域，这些项目涉及细胞，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），细菌蛋白质分泌（Lacy，Cover）， 基底膜结构（Weaver，哈德逊）和细胞膜运输（Weaver，Goldenring， Zhou）。这些研究对于我们理解疾病和与之相关的疾病具有重要意义。 出生缺陷、癌症、消化系统疾病和许多其他人类健康问题。Crossbeam 550将在 范德比尔特细胞成像共享资源（CISR）和范德比尔特冷冻EM之间的合作 结构生物学中心（Center for Structural Biology，CSB）。它将由范德比尔特小组管理 映像共享资源。CISR将为引入范德比尔特的研究人员提供基础设施 这些新技术，培训用户操作仪器，并应用既定的商业模式， 支持系统的长期维护。蔡司Crossbeam 550 FIB的收购 SEM将允许生物样品在尺度和空间上的前所未有的体积成像能力。 决议目前还没有提供给范德比尔特社区，并将促进研究的一套不同的 话题