Acquisition of a Glacios Cryo-TEM for the Biological Sciences Imaging Resource

采购 Glacios Cryo-TEM 用于生物科学成像资源

• 批准号: 10632855
• 负责人: SCOTT M STAGG
• 金额: $ 200万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2024-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10632855
• 关键词: 3-Dimensional; Aging; Amyloid; Biological Sciences; Clustered Regularly Interspaced Short Palindromic Repeats; Computer software; Contract Services; Cryoelectron Microscopy; Data Collection; Electron Microscope; Electrons; Filament; Florida; Freezing; Funding; Health; Hydration status; Image; Marketing; Methods; Microscope; Minor; Modification; Molecular Weight; Muscle; Muscle function; Myocardium; Preparation; Request for Applications; Research; Resolution; Resources; Ribosomes; Robotics; Running; Sampling; Specimen; Structure; Sulfur Metabolism Pathway; Time; Titan; United States National Institutes of Health; Universities; Vesicle; detector; electron tomography; field emission gun; instrument; particle; reconstruction; repaired; screening; structural biology; trafficking; transmission process

PROJECT SUMMARY The Biological Science Imaging Resource (BSIR) at Florida State University is the focal point for cryogenic electron microscopy (cryo-EM) in the state of Florida and the Southeast. No other University facility in the region has a comparable emphasis or facilities. This application requests funds to replace an ageing conventional transmission electron microscope (TEM) called a CM120 that was built in 1997 is no longer supported by service contract and no longer has parts available for repair. We will replace the CM120 with a ThermoFisher Glacios cryo-EM. The Glacios will serve two purposes: efficient specimen screening and high-resolution data collection, especially for low molecular weight samples. The Glacios will support the needs of 5 major users and 2 minor users. The Glacios is a 200 keV TEM with a field emission gun (FEG). It is equipped with an autoloader that can robotically load up to 12 samples at a time. The autoloader specimen carriers (cartridges) are compatible with the existing high-end Titan Krios at FSU making the Glacios and ideal screening microscope. The Glacios will be equipped with a Direct Electron (DE) Apollo electron detector. The Apollo will be the primary data collection camera for the microscope. We have the first commercial Apollo equipped on our Titan Krios, and it has produced the highest resolution 3D reconstruction at FSU. The Apollo is the fastest detector on the market which makes it ideal for screening. The combination of the FEG and the Apollo make the Glacios a good data collection instrument as well as a screening instrument. The study of frozen-hydrated specimens by electron tomography and single particle methods is a large and growing research emphasis worldwide. The robotic capabilities of the Titan-Krios with its ability to run state of the art automated EM software, such as Leginon, are well adapted to this need, but ultimately, efficient utilization of the Titan Krios requires preparation of quality specimens that have been prescreened. The Glacios will support NIH and NSF funded research of the major users which encompasses the atomic resolution imaging of muscle filaments, which has health relevence to cardiac muscle function; ribosome modification and CRISPR structure & function, the mechanisms of vesicle trafficking sulfur metabolism, amyloid structural biology, and automated structure determination.

项目摘要 佛罗里达州立大学的生物科学成像资源（BSIR）是低温的焦点 佛罗里达州和东南部的电子显微镜（冷冻EM）。没有其他大学的设施 地区具有可比的重点或设施。该申请要求资金更换老化 1997年建造的常规传输电子显微镜（TEM）称为CM120 由服务合同支持，不再可以维修零件。我们将用 热泡晶状体冷冻EM。冰川将有两个目的：有效的标本筛选和 高分辨率数据收集，特别是对于低分子量样品。冰川将支持 5个主要用户和2个次要用户的需求。 Glacios是带有场排放枪（FEG）的200 kev tem。 它配备了一个自动加载器，该自动加载器一次可以一次装载多达12个样品。自动加载器 标本载体（墨盒）与FSU现有的高端泰坦krios兼容 冰川和理想筛选显微镜。冰川将配备直接电子（DE）Apollo 电子检测器。 Apollo将是显微镜的主要数据收集摄像头。我们有 第一个商业阿波罗（Apollo FSU的重建。阿波罗是市场上最快的检测器，它是筛查的理想选择。 FEG和Apollo的结合使冰川成为一个很好的数据收集工具以及 筛选工具。电子断层扫描和单个粒子对冷冻水合的标本的研究 方法是全球范围内越来越多的研究重点。 Titan-Krios的机器人能力 凭借其运行最先进的自动化EM软件（例如Leginon）的能力，可以很好地适应这种需求， 但是最终，泰坦krio的有效利用需要准备已经 预先筛选。 Glacios将支持NIH和NSF资助的主要用户研究 涵盖了肌肉丝的原子分辨率成像，该成像与心脏具有健康相关性 肌肉功能；核糖体修饰和CRISPR结构和功能，囊泡的机制 运输硫代谢，淀粉样蛋白结构生物学和自动结构确定。