Thermo Scientific Glacios cryo-TEM

项目摘要 我们建议购买一台Thermo Scientific Glacios，一台200 kV低温透射电子显微镜， 利用冷冻电镜的生长技术来推进我们的生物学研究。冰川将是 Cryo-EM设施的一部分，结构生物学中心的共享资源。我们 研究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。冰川的增加将是必不可少的， 我们的研究人员将进一步推进他们的研究，超越目前使用我们的 现有文书。具体而言，冰川将是必不可少的：（1）低层构造分析 利用200 kV加速电压的分子量蛋白质，（2）使用MicroED方法， 微晶的电子衍射，（3）大分子复合物的低温原位研究， 电子断层扫描，和（4）标本优化，以提高我们的效率，以解决在高- 高分辨精细构象的品种和小分子，如配体、药物和离子。的 Glacios提供的技术对于打破NIH资助的研究的新局面至关重要 主要用户的项目，包括突触传递和可塑性研究（中川）， 原位突触结构和突触囊泡融合（Zhou），艰难梭菌毒素病理学 （Lacy），核mRNA转运（Ren），DNA复制和修复（Eichmann），钙信号传导 （Karakas）、原位病毒组装和结构（Wan）、囊泡和膜运输 （杰克逊）和细菌信号传导（艾弗森）。此外，11个小用户计划利用冰川， 研究膜蛋白功能、炎症信号传导、基因组中的关键生物学问题 维持、细菌-宿主相互作用、离子通道介导的信号传导、脂质信号传导和 生物合成和分子马达。将使用冰川的研究项目的产出将 实质性地推进我们对人类生理学，病理学， 和疾病治疗学。Glacios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， 相关的科学兴趣。事实上，冰川将被安置在工程和科学大楼， 它的目的是通过一个三层楼的，综合的， 协作实验室空间连接医学院，艺术和科学学院， 工程学院。

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.