Thermo Scientific Glacios cryo-TEM

Thermo Scientific Glacios 冷冻透射电镜

• 批准号: 10175401
• 负责人: Terunaga Nakagawa
• 金额: $ 200万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-15 至 2022-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10175401
• 关键词: Acceleration; Anabolism; Architecture; Arts; Bacteria; Biological; Biomedical Engineering; Calcium Signaling; Cells; Chemistry; Clostridium difficile; Collaborations; Cryo-electron tomography; Cryoelectron Microscopy; Crystallization; DNA Repair; DNA biosynthesis; Discipline; Disease; Electron Microscope; Engineering; Floor; Funding; Genome; Human; In Situ; Inflammatory; Investigation; Ion Channel; Ions; Laboratories; Ligands; Link; Lipids; Macromolecular Complexes; Maintenance; Mediating; Membrane; Membrane Proteins; Messenger RNA; Micro Electron Diffraction; Minor; Molecular; Molecular Conformation; Molecular Motors; Molecular Weight; Nuclear; Output; Pathology; Pharmaceutical Preparations; Physiology; Proteins; Records; Research; Research Personnel; Research Project Grants; Resolution; Resource Sharing; Schools; Science; Signal Transduction; Specimen; Structural Biologist; Structure; Synapses; Synaptic Transmission; Synaptic Vesicles; Synaptic plasticity; Technology; Therapeutic; Toxin; United States National Institutes of Health; Vesicle; Virus Assembly; college; design; instrument; interest; medical schools; novel; particle; protein function; small molecule; structural biology; trafficking; transmission process; voltage

PROJECT SUMMARY We propose to acquire a Thermo Scientific Glacios, a 200kV cryo-transmission electron microscope, to advance our biological investigation using the growing technologies of cryo-EM. The Glacios will be a part of the Cryo-EM Facility, a shared resource of the Center for Structural Biology. Our investigators have established records of using single particle analysis and discovering novel biological molecular mechanisms at the atomic level. The addition of the Glacios will be essential for our investigators to further advance their research beyond what is currently possible using our existing instruments. Specifically, the Glacios will be essential for: (1) structural analyses of low molecular weight proteins utilizing 200kV acceleration voltage, (2) the MicroED approach that uses electron diffraction of micro-crystals, (3) investigation of macromolecular complexes in situ using cryo- electron tomography, and (4) specimen optimization to advance our efficiency to resolve at high- resolution fine conformational varieties and small molecules, such as ligands, drugs, and ions. The technology provided by the Glacios is essential for breaking new ground on the NIH-funded research projects of the major users, which include studies of synaptic transmission and plasticity (Nakagawa), in situ synapse architecture and synaptic vesicle fusion (Zhou), Clostridioides difficile toxin pathology (Lacy), nuclear mRNA transport (Ren), DNA replication and repair (Eichmann), calcium signaling (Karakas), in situ viral assembly and architectures (Wan), vesicle and membrane trafficking (Jackson), and bacterial signaling (Iverson). Additionally, 11 minor users plan to exploit the Glacios to investigate critical biological questions in membrane protein function, inflammatory signaling, genome maintenance, bacteria-host interactions, ion channel mediated signaling, lipid signaling and biosynthesis, and molecular motors. The outputs of the research projects that will use the Glacios will substantially advance our basic understanding on the mechanisms of human physiology, pathology, and disease therapeutics. The Glacios will initially be used by structural biologists and cell biologists, with the userbase predicted to extend into the fields of chemistry and bioengineering who have related scientific interests. In fact, the Glacios will be housed in the Engineering and Science Building, which is designed to nurture collaborations between disciplines via a three-floor, integrative, and collaborative laboratory space linking the School of Medicine, College of Arts and Sciences, and the School of Engineering.

项目总结 我们计划购买一台Thermo Science Glacios，一台200千伏低温透射式电子显微镜， 利用低温EM不断发展的技术来推进我们的生物研究。冰川将会是 冷冻-EM设施的一部分，结构生物学中心的共享资源。我们的 调查人员已经建立了使用单粒子分析和发现新的 原子水平上的生物分子机制。冰川的加入将是至关重要的 我们的研究人员将进一步推进他们的研究，使他们的研究超越目前使用我们的 现有的工具。具体地说，冰川对于：(1)低气压的结构分析将是必不可少的。 利用200KV加速电压的分子量蛋白质，(2)使用 微晶的电子衍射；(3)低温原位研究大分子络合物。 电子断层扫描，以及(4)样品优化，以提高我们的效率，以解决在高- 拆分精细的构象变化和小分子，如配体、药物和离子。这个 冰川提供的技术对于在NIH资助的研究中开辟新天地至关重要 主要使用者的项目，包括突触传递和可塑性的研究(Nakagawa)， 原位突触构筑和突触囊泡融合(周)艰难梭菌毒素病理学 (Lacy)、核mRNA转运(REN)、DNA复制与修复(Eichmann)、钙信号转导 (Karakas)、病毒原位组装和结构(WAN)、囊泡和膜运输 (杰克逊)和细菌信号(艾弗森)。此外，11名未成年用户计划利用冰川 研究膜蛋白功能、炎症信号、基因组等关键生物学问题 维持，细菌-宿主相互作用，离子通道介导的信号，脂质信号和 生物合成和分子马达。将使用冰川的研究项目的成果将 极大地提高了我们对人类生理、病理、 和疾病治疗学。冰川最初将由结构生物学家和细胞生物学家使用， 预计用户群将扩展到化学和生物工程领域，这些领域 相关的科学兴趣。事实上，冰川将被安置在工程和科学大楼， 旨在通过三层、集成和 合作实验室空间，连接医学院、文理学院和 工程学院。