Chameleon Sample Preparation Device for Cryo-EM

用于冷冻电镜的变色龙样品制备装置

• 批准号: 10440804
• 负责人: Andres Leschziner
• 金额: $ 42.04万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10440804
• 关键词: Address; Area; Automation; Biogenesis; California; Cryoelectron Microscopy; DNA Repair; DNA biosynthesis; Devices; Electron Microscope; Electron Microscopy; Funding; Image Analysis; Immunology; Institution; Manuals; Membrane; Nerve Degeneration; Preparation; Process; Proteins; Reproducibility; Resolution; Sampling; Specimen; Structure; Techniques; Technology; Training; United States National Institutes of Health; Universities; cryogenics; electron optics; improved; innovation; instrument; novel strategies; particle; trafficking; virology

PROJECT SUMMARY Recent technological advances in cryogenic-electron microscopy (cryo-EM) have heralded a “resolution revolution” that has made it possible for high-resolution structures to be determined for a variety of targets that had eluded other structural techniques. Despite these advances, cryo-EM grid preparation remains a formidable roadblock to successful structure determination in many cases. Indeed, despite improvements in electron optics, camera technology, and image analysis, and a general drive towards automation, cryo-EM grid preparation remains a laborious and unpredictable process that requires extensive manual user input for optimization. The University of California, San Diego (UCSD) has established a world-class cryo-EM facility with state-of-the-art electron microscopes and staff, yet numerous projects across campus remain stalled due to an inability to prepare high-quality cryo-EM grids from purified specimens. To address this bottleneck in the cryo-EM workflow at UCSD, we propose to purchase an SPT Labtech Chameleon sample vitrification instrument. This instrument represents the first major innovation in cryo-EM grid preparation in over 30 years, with an entirely novel approach to sample application that yields more reproducible cryo-EM grids, improves particle quality and distribution, and generally increases the likelihood of high-resolution structure determination. The instrument will be centrally housed and operated within the recently renovated UCSD Cryo-EM Facility and will be available to all trained users at UCSD, surrounding institutions in the La Jolla “Mesa” area, and across California. For users at UCSD, this instrument will catalyze progress in a broad range of NIH-funded, biomedically-important projects spanning neurodegeneration, virology, membrane trafficking, DNA replication and repair, immunology, and protein biogenesis and folding.

项目概要 低温电子显微镜 (cryo-EM) 的最新技术进步预示着“分辨率 “革命”使得确定各种目标的高分辨率结构成为可能 尽管有这些进步，冷冻电镜网格的制备仍然是一项艰巨的任务。 事实上，尽管电子光学有所改进，但在许多情况下仍是成功确定结构的障碍。 相机技术、图像分析以及自动化、冷冻电镜网格准备的总体推动 仍然是一个费力且不可预测的过程，需要大量的手动用户输入来进行优化。 加州大学圣地亚哥分校 (UCSD) 建立了世界一流的冷冻电镜设施，配备最先进的设备 电子显微镜和工作人员，但校园内的许多项目由于无法 从纯化的样本中制备高质量的冷冻电镜网格，以解决冷冻电镜工作流程中的这一瓶颈。 在 UCSD，我们建议购买 SPT Labtech Chameleon 样品玻璃化仪器。 代表了 30 多年来冷冻电镜网格制备领域的首次重大创新，采用了全新的方法 样品应用可产生更具可重复性的冷冻电镜网格，改善颗粒质量和分布，以及 通常会增加仪器集中测定高分辨率结构的可能性。 位于最近翻新的加州大学圣地亚哥分校冷冻电镜设施内并运行，所有接受过培训的人员都可以使用 加州大学圣地亚哥分校、拉霍亚“梅萨”地区周边机构以及整个加州的用户 对于加州大学圣地亚哥分校的用户， 该工具将促进 NIH 资助的广泛生物医学重要项目的进展，这些项目涵盖 神经变性、病毒学、膜运输、DNA 复制和修复、免疫学和蛋白质 生物发生和折叠。