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) 是一种用于确定结构的强大技术 来自冷冻水合样本的大分子复合物，绕过了其他物质施加的限制 结构生物学技术，如 X 射线晶体学和 NMR。冷冻电镜依赖于个体成像 高放大倍数的分子，然后将数十万个分子副本的图像平均在一起 分子在三个维度上。冷冻电镜在结构生物学方法中的独特之处在于 能够从混合的分子群中解析不同的构象状态，使其成为理想的 在接近生理条件下研究柔性和异质大分子。 冷冻电镜领域的最新技术进步已经在可实现的方面产生了巨大的飞跃 分辨率，近原子分辨率结构已成为常规。这意味着冷冻电镜已经到来 在可确定结构的准确性方面可与 X 射线晶体学相媲美，并扩展 可以进行高分辨率结构分析的大分子类型。这些 一些研究人员认为，结构生物学领域的进步已经发生了结构性转变 大规模放弃其他结构技术，而其他人则急于采用新技术 对现有方法的补充。两项技术发展推动了这一转变： 新一代电子显微镜专为冷冻电镜的独特限制而设计，并经过改进 光学和更高的吞吐量，以及先进的直接电子探测器，可显着提高 记录图像的分辨率。华盛顿大学已经投资了直接探测器技术，我们在这里 请求资金购买高性能冷冻电子显微镜 FEI Talos Arctica。 Talos Arctica 是一款高性能电子显微镜，其设计初衷是为了 用于冷冻电镜。改进包括恒定功率物镜，以提高热稳定性 并减少滞后，用于自动化和无污染样本传输的低温自动装载机， 更好的真空系统，用于保持样品不受污染，压电平台用于改进 机械精度和稳定性，以及更好的环境控制和封闭平台 稳定。 Talos Arctica 所体现的改进的净效果是提高了 获取更高质量的数据。显微镜的质量、可靠性和通量将扩大 整个校园和该地区的用户都可以使用冷冻电镜。该显微镜将能够 新的研究方向，并补充基础等不同领域正在进行的研究项目 细胞生物学与生物化学、传染病与疫苗开发、膜蛋白 结构、细胞应激反应、新陈代谢、神经生物学和癌症。