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.

项目摘要 低温电子显微镜（Cryo-EM）的最新技术进步预示了“分辨率 革命”，这使得高分辨率结构有可能确定各种目标 已经避免了其他结构技术。尽管有这些进展，但冷冻EM网格的准备仍然是强大的 在许多情况下，成功确定结构的障碍。确实，电子光学的目的地改进， 摄像机技术和图像分析，以及朝着自动化的一般驱动器，冷冻EM电网准备 仍然是实验室和不可预测的过程，需要大量的手动用户输入才能优化。这 加利福尼亚大学圣地亚哥大学（UCSD）已建立了一个世界级的低温EM设施 电子显微镜和员工，但是由于无法 从纯化的标本中制备高质量的低温EM网格。在Cryo-EM工作流程中解决此瓶颈 在UCSD，我们建议购买SPT Labtech变色龙样品玻璃化仪器。这个乐器 代表了30多年来Cryo-Em Grid制备中的第一个主要创新，采用了一种完全新颖的方法 为了采样的应用，该应用产生更多可再现的冷冻网格，改善粒子质量和分布，以及 通常会增加高分辨率结构确定的可能性。该乐器将集中 在最近经过翻新的UCSD Cryo-EM设施中安装和操作，所有受过培训的 UCSD的用户，La Jolla“ Mesa”地区以及整个加利福尼亚州的机构。对于UCSD的用户， 该工具将在广泛的NIH资助，生物医学重要的项目中催化进步 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。