Acquisition of a high resolution, high throughput cryo-electron microscope

购置高分辨率、高通量冷冻电子显微镜

• 批准号: 9273775
• 负责人: Justin M Kollman
• 金额: $ 200万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9273775
• 关键词: Adopted; Biochemistry; Bypass; Cellular Stress Response; Cellular biology; Communicable Diseases; Complement; Cryoelectron Microscopy; Data Quality; Development; Dimensions; Electron Microscope; Electrons; Freezing; Funding; Generations; Hydration status; Image; Individual; Macromolecular Complexes; Malignant Neoplasms; Mechanics; Membrane Proteins; Metabolism; Microscope; Molecular Conformation; Molecular Structure; Neurobiology; Optics; Performance; Physiological; Population; Research; Research Personnel; Resolution; Specimen; Structure; System; Techniques; Technology; Vacuum; X-Ray Crystallography; design; detector; flexibility; improved; lens; macromolecule; new technology; programs; protein structure; quantum; structural biology; vaccine development

Project Abstract Cryo-electron microscopy (cryo-EM) is a powerful technique for determining the structures of macromolecular complexes from frozen hydrated specimens, bypassing constraints imposed by other structural biology techniques like X-ray crystallography and NMR. Cryo-EM relies on imaging individual molecules at high magnification, then averaging together images of hundreds of thousands of copies of the molecule in three dimensions. Cryo-EM is unique among structural biology approaches in being able to resolve different conformational states from a mixed population of molecules, making it ideal to study flexible and heterogeneous macromolecules under near-physiological conditions. Recent technological advances in cryo-EM have generated a quantum leap in achievable resolution, with near-atomic resolution structures becoming routine. This means that cryo-EM has come to rival X-ray crystallography in the accuracy of structures that can be determined, as well as expanding the types of macromolecules that can be subjected to high-resolution structural analysis. These advances have created tectonic shifts in the landscape of structural biology, with some investigators abandoning other structural techniques wholesale and others rushing to adopt the new technologies as a compliment to existing approaches. Two technological developments have driven this shift: a new generation of electron microscopes designed for the unique constraints of cryo-EM with improved optics and higher throughput, and advanced direct electron detectors that dramatically improve the resolution of recorded images. UW has already invested in direct detector technology, and here we are requesting funds to acquire a high performance cryo-electron microscope, the FEI Talos Arctica. The Talos Arctica is a high performance electron microscope designed from the ground up to be used for cryo-EM. Improvements include a constant power objective lens for greater thermal stability and reduced hysteresis, a cryo-autoloader for automated and contamination free specimen transfer, better vacuum system for maintaining specimens without contamination, a piezo stage for improved mechanical precision and stability, and an enclosed platform for better environmental control and stability. The net effect of the improvements embodied in the Talos Arctica is an increased rate of acquisition of higher quality data. The quality, reliability, and throughput of the microscope will broaden the accessibility of cryo-EM for users across campus and in the region. This microscope will enable new lines of research and complement ongoing research programs in fields as diverse as fundamental cell biology and biochemistry, infectious diseases and vaccine development, membrane protein structure, cellular stress responses, metabolism, neurobiology, and cancer.

项目摘要 冷冻电子显微镜（Cryo-EM）是确定结构的强大技术 来自冷冻水合标本的大分子复合物，绕过其他施加的约束 X射线晶体学和NMR等结构生物学技术。冷冻EM依赖成像个体 高放大倍数的分子，然后将数十万份的图像平均 分子在三个维度上。 Cryo-Em在结构生物学方法中是独一无二的 能够从混杂的分子种群中解析不同构象状态，使其理想 在近乎生理的条件下研究柔性和异质大分子。 Cryo-EM的最新技术进步已经在可实现的方面产生了量子飞跃 分辨率，近乎原子的分辨率结构成为常规。这意味着冷冻EM来了 以可以确定的结构的准确性以及扩展 可以进行高分辨率结构分析的大分子类型。这些 进步在结构生物学的景观中创造了构造转变，一些研究者 放弃其他结构技术批发和其他急于采用新技术作为 对现有方法的称赞。两个技术发展驱动了这一转变：一个新的 具有改进的Cryo-EM独特约束的电子显微镜的生成 光学和较高的吞吐量和高级直接电子检测器，这些检测器极大地改善了 记录图像的分辨率。 UW已经投资了直接检测器技术，我们在这里 要求资金获得高性能冷冻电子显微镜，Fei Talos Arctica。 Talos Arctica是从地面设计的高性能电子显微镜 用于冷冻EM。改进包括恒定的功率目标镜头，以实现更大的热稳定性 并减少了滞后，是一种用于自动化和无污染的标本转移的冷冻载体加载器， 更好的真空系统，用于维持没有污染的标本，一个压电阶段以改进 机械精度和稳定性，以及一个封闭的平台，用于更好的环境控制和 稳定。塔洛斯arctica体现的改进的净效应是增加的速度 获取更高质量的数据。显微镜的质量，可靠性和吞吐量将扩大 校园和该地区用户的冷冻EM可访问性。该显微镜将启用 在类似基本的领域中，新的研究和补充正在进行的研究计划 细胞生物学和生物化学，传染病和疫苗发育，膜蛋白 结构，细胞应激反应，代谢，神经生物学和癌症。