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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

基本信息

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
项目类别：
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

项目摘要

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.

项目总结