TRANSMISSION ELECTRON MICROSCOPE

这份申请是由一组研究人员提交的，他们建议为NIH购买透射电子显微镜（TEM）。资助的研究项目。 所有人都在使用TEM来回答基础分子、细胞和发育生物学中的重要问题。我们建议用具有数字成像能力的现代“主力”仪器替换制造商不再支持的25岁TEM（Philips 301），并且不再有许多可用的替换部件。在考虑了许多选择之后，我们选择了配备Advantage R数字CCD相机系统的Philips/FEI 208 S显微镜，因为它具有多功能性，成本低，以及Philips在该地理区域提供的出色技术服务。4个主要用户实验室和4个次要用户实验室占9个NIH -和1个N. S.F. -资助的研究项目涵盖广泛的主题，所有这些都大大受益于TEM和免疫电子显微镜的分析。该仪器的主要用户所涵盖的主题包括哺乳动物细胞分泌和内吞途径中的膜运输（Brown博士），极化上皮细胞和酵母酿酒酵母中肌动蛋白细胞骨架的结构和功能（Bretscher博士），微管在S. cerevesiae（Huffaker博士），以及Rous肉瘤病毒的组装和包装（沃格特博士）。其他研究课题也受益于TEM分析，但在不太一致的基础上，因此形成了一个小的用户组，包括非洲爪蟾和酵母中有丝分裂检查点对中期到后期转换的调节（陈博士），核纤层蛋白Ya的作用，这是从减数分裂到有丝分裂的过渡在受精的果蝇卵必不可少的（Dr. Wolfner），C.线虫（Kemphues博士），以及胆固醇在膜结构中的作用（Feigenson博士）。这个小组的成员将继续利用TEM在他们的研究，事实上，我们预测，在“后基因组测序”时代，越来越多的项目将需要EM和免疫电子显微镜分析。

This application is being submitted by a group of investigators who propose to purchase a transmission electron microscope (TEM) for their NIH.-funded research programs. All are using TEM to answer important questions in basic molecular, cellular, and developmental biology. We propose to replace a 25 year old TEM (Philips 301) that is no longer going to be supported by the manufacturer, and for which there are no longer many replacement parts available, with a modern "workhorse" instrument that has digital imaging capacity. After considering many options, we have chosen a Philips/FEI 208S microscope with Advantage R Digital CCD camera system because of its versatility, low cost, and the outstanding technical service provided by Philips in this geographic region. The 4 major and 4 minor user laboratories account for 9 NIH - and l N.S.F.-funded research projects that cover a wide range of topics, all of which greatly benefit from analysis by TEM and immunoelectron microscopy. The topics covered by the major users of this instrument include membrane trafficking in the secretory and endocytic pathways of mammalian cells (Dr. Brown), the structure and function of the actin cytoskeleton in polarized epithelial cells and in the yeast Saccharomyces cerevesiae (Dr. Bretscher), the role of microtubules in chromosome segregration in S. cerevesiae (Dr. Huffaker), and the assembly and packaging of Rous sarcoma virus (Dr. Vogt). Other research topics that also benefit from TEM analysis, but on a less consistent basis and therefore forming a minor user group, include the regulation of metaphase-to-anaphase transition by mitotic checkpoints in Xenopus and yeast (Dr. Chen), the role of the nuclear lamina protein Ya that is essential for the transition from meiosis to mitosis in the fertilized Drosophila egg (Dr. Wolfner), establishment of polarity in the early embryo of C. elegans (Dr. Kemphues), and the role cholesterol in membrane structure (Dr. Feigenson). The members of this group will continue to utilize TEM in their research, and, in fact, we predict that in the "post-genome sequencing" era, an increasing number of projects will require analyses by EM and immunoelectron microscopy.