120kV Transmission Electron Microscope for Core Electron Microscopy Facility

项目总结/摘要 该提案要求提供资金，用于购买JEOL JEM-1400型透射电子显微镜 具有低温功能，可以用数码相机取代我们已有33年历史的模拟JEOL JEM-100 CX II TEM 位于北达科他州州立大学（NDSU）的电子显微镜中心核心设施中。这个州- 最先进的仪器将扩大NDSU一些NIH资助的研究人员的研究能力。它 将为TEM提供可靠的常规访问，该TEM能够记录低温保存样品的图像 （Cryo-EM），并继续提供可靠的负染色能力。TEM和提供的 支持附件将允许研究者常规制备、筛选和表征样本， 被“冻结”了。因此，NDSU研究人员将能够扩展到涉及低温的研究领域， EM单粒子重建、Cryo-EM层析成像和Cryo-EM微晶电子衍射。的 所要求的仪器能够筛选所有这些技术的样本和数据收集。这 结构数据为理解蛋白质结构-功能关系提供了关键信息， 化学结构，软聚合物结构，以及控制生物医学相关的整体细胞排列 流程.目前，NDSU没有这种仪器，并且在一个区域内没有Cryo-EM设施。 常规旅行的合理距离：距离我们机构最近的两个cryoTEM位于240英里/4处 750英里/11小时的驾驶时间。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，并且对于 investigators.访问该仪器将提供进入NIH高分辨率冷冻电子 通过允许常规访问收集初步数据所需的筛选范围， 要充分利用全国服务中心。作为区域内符合IDEA资格的机构，请说明 资源可以对NDSU正在进行的科学类型产生变革性影响。仪器 将被安置在由两名全职科学家管理的核心设施中，由NDSU提供支持。因此，获取和 仪器的维护是在知识渊博的专家的直接监督下进行的， 仪器的寿命。将被解决的直接相关的项目涉及1）蛋白质：蛋白质 参与细菌细胞表面信号传导的相互作用，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G-蛋白偶联受体， 自噬调节，和4）用于药物递送的纳米颗粒组装体。总的来说，我们的用户群代表了 三所学院五个系的调查人员，该仪器将促进以下领域的研究： 与药理学和材料有关的。NDSU的Cryo-EM仪器的存在将大大提高 这些和其他NIH相关项目的研究方向。它将大大增加学生的培训 机会和教师的研究生产力。

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.