Transmission Electron Microscope

描述（由申请人提供）：本申请要求资助购买高分辨率日立H-7650-II透射电子显微镜（TEM）和断层扫描和数字成像配件。H-7650-II将被安置在康涅狄格大学健康中心（UCHC）的中央电子显微镜设施（CEMF）中，这是一个由健康中心研究咨询委员会，校外赠款和用户费用支持的中央研究设施。新的TEM将取代其中一个CEMF两个旧的和不可靠的TEM，提供目前在设施中不存在的能力。H-7650-II的运作将由PI和Co-Is管理，并由现有的教师咨询委员会监督。要求的配件包括AMT XR-611侧装和Advantage ERB底装CCD相机、用于断层扫描的自动倾斜图像采集系统和用于3D重建的Hitachi软件、冷指、轨迹球载物台控制、多样本保持器、水再循环器和备用Wehnelt。一个由15名NIH资助的科学家组成的主要用户组，研究超分子复合物、细胞器、细胞和组织的结构和功能，将占仪器使用量的约80%。需要断层扫描分析的项目包括鞭毛动力蛋白复合物的结构分析，单纯疱疹病毒门脉环的分析，细胞内纳米制造结构的表征，三维肌肉肌节结构与二次谐波图像的相关性，细菌细胞骨架的膜附着位点的分析，以及外分泌细胞中高尔基复合物的结构。有几个项目将研究实验操作或基因改变小鼠神经系统区域的结构变化。其他项目将采用薄切片的免疫金标记、细菌皮利的负染色免疫标记和磷酸钙晶体的电子衍射分析。如果时间允许，该仪器将提供给UCHC的其他科学家。拟议的项目与许多人类疾病有关，包括早期发育缺陷、癫痫和多发性硬化症等神经系统疾病、血管疾病、膀胱功能障碍、颅面区域遗传性疾病和细菌感染。数码相机将促进快速图像采集和分析，H-7650-II的自动化功能使其成为多用户设施的理想仪器。新的TEM还将通过电子显微镜研究生课程和实验室轮换和研究奖学金期间的实践经验，为学生和年轻科学家提供更多的培训机会。UCHC行政部门致力于为其教师提供最先进的研究设施。这表现在它承诺提供仪器的部分购买费用，购买仪器维护合同，并通过购买辅助制备仪器进一步升级CEMF。公共卫生相关性：细胞和组织成分的结构分析提供了有关正常功能和病理状况的重要信息。本申请中要求的高分辨率透射电子显微镜和配件将在很大程度上用于调查几种疾病相关现象的研究项目。这些项目包括与神经系统和心血管疾病、癌症、遗传性疾病、衰老疾病和传染病有关的研究。

DESCRIPTION (provided by applicant): This application requests funding to purchase a high-resolution Hitachi H-7650-II transmission electron microscope (TEM) and accessories for tomography and digital imaging. The H-7650-II will be housed in the Central Electron Microscope Facility (CEMF) at the University of Connecticut Health Center (UCHC), a central research facility supported by the Health Center Research Advisory Committee, extramural grants and user fees. The new TEM will replace one of the CEMFs two old and unreliable TEMs, providing capabilities not currently present in the facility. The operation of the H-7650-II will be managed by the PI and Co-Is, with oversight by an existing faculty advisory committee. The requested accessories include AMT XR-611 side mount and Advantage ERB bottom mount CCD cameras, an automatic tilt image acquisition system for tomography and Hitachi software for 3D reconstructions, cold finger, trackball stage control, multi-specimen holder, water recirculator and spare Wehnelt. A major user group of 15 NIH-funded scientists investigating the structure and function of supramolecular complexes, organelles, cells and tissues will account for approximately 80% of the instrument's use. Projects requiring tomographic analysis include structural analyses of flagellar dynein complexes, analysis of Herpes simplex virus portal rings, characterization of intracellular nanofabricated structures, correlation of 3D muscle sarcomere structure with 2nd harmonic images, analysis of membrane attachment sites of the bacterial cytoskeleton, and Golgi complex structure in exocrine secretory cells. Several projects will examine structural changes in regions of the nervous system of experimentally manipulated or genetically altered mice. Other projects will employ immunogold labeling of thin sections, negative stain immunolabeling of bacterial pili, and electron diffraction analysis of calcium phosphate crystals. As time permits, the instrument will be made available to other UCHC scientists. The proposed projects have relevance to a number of human conditions including early developmental defects, nervous system disorders such as epilepsy and multiple sclerosis, vascular disease, bladder dysfunction, inherited disorders of the craniofacial region, and bacterial infections. The digital cameras will facilitate rapid image acquisition and analysis, and the automated features of the H-7650-II make it an ideal instrument for a multiuser facility. The new TEM also will provide increased opportunities for training of students and young scientists through a graduate course in electron microscopy and hands- on experience during laboratory rotations and research fellowships. The UCHC administration is committed to providing state-of-the-art research facilities for its faculty. This is demonstrated by its commitment to provide a portion of the purchase cost of the instrument, to purchase instrument maintenance contracts, and to further upgrade the CEMF through the acquisition of ancillary preparative instruments. Public Health Relevance: Project Narrative Structural analyses of cellular and tissue components provide important information regarding normal function and pathological conditions. The high-resolution transmission electron microscope and accessories requested in this application will be used to a great extent in research projects investigating several disease-related phenomena. These projects include studies relevant to nervous system and cardiovascular diseases, cancer, inherited diseases, diseases of aging, and infectious diseases.