INTEGRATED EM-IMMUNOLABELING INSTRUMENTATION

十个主要用户申请资金，以提供综合仪器，以提供在光学和电子显微镜水平上进行最先进的冷冻固定和免疫标记的能力。这些仪器包括一台带有FSC冷冻附件的徕卡Ultraut UCT切片机。DIATOME钻石刀具、配置用于电子显微镜的Edwards AUTO 306高真空沉积系统、Bal-Tec Jet冷冻装置和Bal-Tec冷冻替代装置。该仪器将被放置在由细胞和结构生物学系和癌症中心共同赞助的中央EM设施中。他们将为设备提供翻新的空间，聘请荷兰乌得勒支大学的Jan Slot博士为我们已经熟练掌握显微手术的科学家提供培训，并提供仪器的持续护理。这10名研究人员正在解决各种生物学问题，每个人都将受益于通过应用快速冷冻固定技术来提高他们的光学和电子显微镜的质量。他们的项目都需要在EM水平上进行免疫定位，而准备和标记Tokuyasu超薄低温切片的能力将加强他们由NIH资助的研究。重要的是，这种仪器将促进健康科学中心许多其他研究人员的研究，这些研究人员需要EM免疫标记，并使用保存良好的细胞和组织来使用各种形态技术进行研究。高真空沉积系统是制备栅格和涂层/阴影分子和大分子络合物的必要系统，以便在电子显微镜下获得最佳的可视化效果。健康科学中心目前没有类似的仪器。这个SGI上的科学家小组有过合作互动的历史，这一背景很好地证明了所请求的仪器的成功共享。

Ten major users request funds to provide integrated instrumentation that will provide the ability to carry out state-of-the-art cryo-fixation and immunounolabeling at both the level of light and electron microscopy. These instruments include a Leica Ultracut UCT microtome with an FSC cryo-attachment. Diatome diamond knives, an Edwards auto 306 high vacuum deposition system configured for electron microscopy, a Bal- Tec Jet Freeze device and a Bal-Tec Freeze substitution unit. This instrumentation will be placed in a central EM-facility that is co- sponsored by the Department of Cellular and Structural Biology and the Cancer Center. They will provide renovated space for the equipment, hire Dr. Jan Slot of the University of Utrecht in the Netherlands to provide training to our scientists already skilled in microtomy and provide for the continued care of the instruments. The ten investigators are addressing various biological questions and each would benefit from improving the quality of their light and electron microscopy by application of rapid freeze fixation techniques. Their projects all require immunolocalization at the EM-level and the ability to prepare and label Tokuyasu-ultrathin cryo sections will enhance their NIH funded research. Importantly this instrumentation will facilitate the research of many other investigators in the Health Sciences Center who require EM-immunolabeling and use of well preserved cells and tissues for study using a variety of morphological techniques. The high vacuum deposition system is necessary to prepare grids and coat/shadow molecules and macromolecular complexes for optimal visualization with the electron microscope. Comparable instruments are not currently available at the Health Sciences Center. The group of scientist on this SGI has a history of collegial interactions, a background that argues well for the successful sharing of the instrumentation requested.