Glacios 200 kV cryogenic transmission electron microscope (cryo-TEM)

项目总结 结构生物学在定义生命的分子结构方面发挥了关键作用，导致了基本的 对大分子的功能和生物学的洞察。在我们的俄亥俄山谷财团中，由 辛辛那提大学、辛辛那提儿童医院医学中心、迈阿密大学和 肯塔基州的所有大学都有完善的结构生物学课程，主要关注X射线结晶学。 这为研究生和博士后研究员提供了一个良好的学习环境 解决围绕生物医学重要性的基本问题的现代结构生物学方法 大分子。然而，近年来，低温电子显微镜(Cryo-EM)作为一种 蛋白质和分子复合体的结构分析不可或缺的技术，因为它可以 产生原子分辨结构，而不存在结晶固有的缺陷。因此，我们的财团 结构生物学家一直在积极努力，将这项技术纳入他们的研究计划， 主要是通过合作和尝试获取其他机构的工具。由于这些原因， 在我们的努力下，我们现在有了足够多的调查人员，这有力地证明了将这项技术带到我们地区的合理性。 辛辛那提大学承诺为这一倡议奠定基础，购买一台Talos 用于阴性染色分析的L120C仪器和用于用玻璃化样品准备网格的Vitbot。这些 投资将使用户能够确定生物样本的质量，以便进一步进行冷冻-EM 分析。为了把我们带到下一个水平，我们正在申请NIH的资金，为Glacios 200千伏显微镜提供 猎鹰4号探测器和自动装载机，将安装在该大学新开发的低温电磁设施中 辛辛那提。冰川将使我们能够进一步优化样品，以便在室内进行结构分析，或提供 使用其他机构或NIH财团提供的先进低温电磁仪器的理由 更高分辨率的研究。总的来说，收购Glacios将直接惠及30多家NIH资助的公司 从事心血管、癌症、传染病和神经科学等重点领域的研究。

PROJECT SUMMARY Structural biology has played a key role in defining the molecular architecture of life, leading to fundamental insights into the function and biology of macromolecules. Within our Ohio Valley consortium, consisting of the University of Cincinnati, Cincinnati's Children's Hospital Medical Center, Miami University, and University of Kentucky, all have well-established programs in structural biology, with a major focus on X-ray crystallography. This has provided an outstanding training environment for graduate students and postdoctoral fellows to learn modern structural biology approaches to address fundamental questions around biomedically important macromolecules. However, in recent years, cryogenic electron microscopy (cryo-EM) has emerged as an indispensable technique for structural analysis of proteins and molecular complexes due to the fact that it can produce atom-resolution structures without drawbacks inherent to crystallization. Thus, our consortium structural biologists have been actively working to incorporate this technology into their research programs, primarily through collaboration and attempts to access instruments at other institutions. As a result of these efforts, we now have a critical mass of investigators that strongly justifies bringing this technology to our region. The University of Cincinnati has committed to laying the groundwork for this initiative by purchasing a Talos L120C instrument for negative stain analysis and a Vitrobot to prepare grids with a vitrified sample. These investments will allow users the ability to ascertain the quality of biological samples for further cryo-EM analysis. To take us to the next level, we are requesting NIH funding for a Glacios 200 kV microscope with Falcon 4 detector and autoloader, which will be housed in a newly developing cryo-EM facility at the University of Cincinnati. The Glacios will allow us to further optimize samples for structural analysis in house, or provide the justification for use of advanced cryo-EM instrumentation available at other institutions or NIH consortia for higher resolution studies. Collectively, the acquisition of the Glacios will directly benefit over 30 NIH funded investigators working in the focus areas of Cardiovascular, Cancer, Infectious Disease, and Neuroscience.