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

项目摘要 结构生物学在定义生命的分子结构方面发挥了关键作用， 深入了解大分子的功能和生物学。在我们的俄亥俄州山谷财团中， 辛辛那提大学、辛辛那提儿童医院医疗中心、迈阿密大学和 肯塔基州，都有完善的程序，在结构生物学，主要侧重于X射线晶体学。 这为研究生和博士后提供了良好的学习环境 现代结构生物学方法来解决生物医学上重要的基本问题 大分子然而，近年来，低温电子显微镜（cryo-EM）已经成为一种 蛋白质和分子复合物的结构分析不可或缺的技术，因为它可以 产生原子分辨率结构而没有结晶所固有的缺点。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使用其他机构的工具。由于这些 经过努力，我们现在有了足够数量的调查人员，这有力地证明了将这种技术带到我们地区的合理性。 辛辛那提大学已承诺通过购买Talos为这一举措奠定基础 L120 C仪器进行阴性染色分析，Vitrobot用玻璃化样品制备网格。这些 投资将使用户能够确定生物样品的质量，以进一步冷冻EM 分析.为了让我们更上一层楼，我们正在申请NIH资助一台Glacios 200 kV显微镜， 猎鹰4号探测器和自动装弹机，将被安置在该大学新开发的冷冻EM设施中 来自辛辛那提。Glacios将使我们能够进一步优化内部结构分析的样品，或提供 使用其他机构或NIH联盟提供的先进冷冻EM仪器的理由， 更高分辨率的研究总的来说，收购Glacios将直接使30多个NIH资助的项目受益。 在心血管、癌症、传染病和神经科学等重点领域工作的研究人员。

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.