120kV Transmission Electron Microscope for Core Electron Microscopy Facility

项目摘要/摘要 该提案要求提供资金购买JEOL JEM-1400型透射电子显微镜(TEM) 具有低温能力，用数码相机取代我们已有33年历史的模拟JEOL JEM-100CX II透射电子显微镜 安装在北达科他州州立大学(NDSU)的电子显微镜中心核心设施中。这种状态- 最先进的仪器将扩大国家卫生研究院资助的NDSU研究人员的研究能力。它 将提供对能够记录低温保存样品的图像的透射电子显微镜的可靠的例行访问 (Cryo-EM)除了继续提供可靠的负染能力。电子显微镜，并提供了 辅助附件将允许调查人员例行准备、筛选和表征符合以下条件的样本 已经被“瞬间冻结”了。因此，南加州州立大学的调查人员将能够扩展到涉及低温- EM单粒子重建、Cryo-EM层析和Cryo-EM微晶电子衍射。这个 所要求的仪器能够为所有这些技术筛选样品和收集数据。这 结构数据为理解蛋白质结构-功能关系以及 控制生物医学相关的化学结构、软聚合物结构和整体细胞排列 流程。目前，NDSU没有这种仪器，而且在 日常旅行的合理距离：距离我们机构最近的两台低温TEM位于240英里/4的位置 小时车程和750英里/11小时车程。因此，用户目前依赖于访问Cryo- EM通过与外部用户付费的远程协作。这是低效的，而且成本高得令人望而却步 调查人员。访问这台仪器将为进入NIH高分辨率冷冻电子提供一条途径 显微镜计划，允许常规访问收集初步数据所需的筛查范围 要充分利用好国家服务中心。作为一个地区有资格的理念机构，这一点 资源可以对国家科技大学正在进行的科学类型产生变革性的影响。这台仪器 将被安置在一个核心设施中，由两名全职科学家管理，并得到NDSU的支持。因此，访问和 仪器的维护都是在知识渊博的专家的直接监督下进行的，从而保护了 乐器的使用寿命。将解决的直接相关项目涉及1)蛋白质：蛋白质 细菌细胞表面信号转导的相互作用，2)了解激动剂/拮抗剂对 蛋白质：蛋白质相互作用涉及G蛋白偶联受体，3)蛋白质：蛋白质相互作用涉及 自噬调节，以及4)用于药物输送的纳米颗粒组件。总体而言，我们的用户组代表 三所大学五个系的研究人员，该仪器将促进领域的研究 与药理学有关的材料。NDSU的Cryo-EM仪器的出现将极大地提高 这些项目和其他与NIH相关的项目的研究方向。它将显著增加对学生的培训 教师的机会和研究效率。

PROJECT SUMMARY/ABSTRACT This proposal requests funds to purchase a JEOL JEM-1400 Transmission Electron microscope (TEM) with cryogenic capabilities to replace our 33-year-old analog JEOL JEM-100CX II TEM with a digital camera housed in the Electron Microscopy Center Core Facility at North Dakota State University (NDSU). This state- of-the-art instrument will expand the research capabilities of a number of NIH funded investigators at NDSU. It will provide reliable routine access to a TEM capable of recording images of cryogenically preserved samples (Cryo-EM) in addition to continue to provide reliable negative staining capabilities. The TEM and supplied supporting accessories will allow investigators to routinely prepare, screen, and characterize samples that have be `flash-frozen'. Thus, NDSU investigators will be able to expand into areas of research involving Cryo- EM single particle reconstruction, Cryo-EM tomography, and Cryo-EM microcrystal electron diffraction. The requested instrument is capable of screening samples and data collection for all these techniques. This structural data provides critical information for understanding protein structure-function relationships as well as chemical structure, soft polymer structure, and overall cellular arrangements that control biomedical related processes. Currently, NDSU does not have this instrumentation and there is no Cryo-EM facility within a reasonable distance for routine travel: the two closest cryoTEMs to our institution are located at 240 miles/4 hours drive time and 750 miles/11 hours drive time. Therefore, users are currently relying on access to Cryo- EM via long distance collaboration with external user fees. This is inefficient and prohibitively expensive for investigators. Access to this instrument will provide a gateway into the NIH High Resolution Cryo-Electron Microscopy Program by allowing routine access to a screening scope needed to collect the preliminary data necessary to make full use of the national service centers. As an IDeA eligible institution in an AREA state this resource can have a transformative impact on the type of science being performed at NDSU. The instrument will be housed in a core facility managed by two full time scientists, supported by NDSU. Thus, access and maintenance of the instrument are under the direct supervision knowledgeable experts thus protecting the longevity of the instrument. Projects of immediate relevance that will be addressed involve 1) protein:protein interactions involved in bacterial cell surface signaling, 2) understanding agonist/antagonist influence on protein:protein interactions involve in a G-protein coupled receptor, 3) protein:protein interactions involved in autophagy regulation, and 4) nanoparticle assemblies for drug delivery. Overall, our user group represents investigators in five departments across three colleges and the instrument will facilitate research in areas related to pharmacology to materials. The presence of Cryo-EM instrument at NDSU will greatly enhance the research directions of these and other projects of NIH relevance. It will significantly increase student training opportunities and research productivity of faculty.