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 该提案要求获得资金以购买聚焦离子束扫描电子显微镜(FIB-SEM)，以 支持美国国立卫生研究院资助的范德比尔特大学和范德比尔特大学的不同研究项目 医疗中心。这种显微镜将可以用来三维重建整个细胞体积 足够的空间分辨率来观察细胞器的精细细节。此卷信息对以下各项至关重要 了解和量化细胞内细胞器和大分子复合体的复杂相互作用。这个 添加冷冻台将扩展FIB-SEM的功能，将玻璃化冷冻细胞的离子研磨添加到 允许在原位和通过低温扫描电子显微镜对大分子结构进行冷冻电子层析成像。蔡司的横梁 550是一种最先进的FIB-SEM，具有高性能的离子柱(3 nm分辨率和高达100 nA的 探头电流)，可以高效地进行FIB-SEM断层扫描，并可以与徕卡冷冻台集成 创造出用于低温电子断层扫描的薄层。我们的主要用户将应用FIB-SEM断层扫描技术 美国国立卫生研究院资助的研究项目的新领域，这些项目涵盖了细胞、发育、 上皮生物学、肠道生物学和癌症生物学等。项目包括对细胞骨架动力学的研究 (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)。综上所述，这些研究对于我们理解疾病和与 出生缺陷、癌症、消化障碍和许多其他人类健康问题。550号横梁将是 Vanderbilt细胞成像共享资源(CISR)和Vanderbilt Cryo-EM之间的合作 结构生物学中心(CSB)内的设施(V-CEM)。它将由范德比尔特细胞管理 正在为共享资源创建映像。该研究所将为在范德比尔特引进研究人员提供基础设施 对于这些新技术，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。总而言之，收购蔡司横梁550 FIB- 扫描电子显微镜将使生物样品在尺度和空间上具有前所未有的体积成像能力 范德比尔特社区目前无法获得的决议，将促进对一套不同的 主题。