TRANSMISSION ELECTRON MICROSCOPE

本申请是由一组研究人员提交的，他们打算为他们的国家卫生研究院购买一台透射电子显微镜（TEM）。美国资助的研究项目。所有人都在使用TEM来回答基本分子、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”仪器替换一台已使用25年的TEM (Philips 301)，因为它不再由制造商提供支持，也不再有许多可用的替换部件。在考虑了很多选择后，我们选择了Philips/FEI 208S显微镜与Advantage R数字CCD相机系统，因为它的通用性，低成本，以及飞利浦在这个地理区域提供的出色的技术服务。4个主要和4个次要用户实验室负责9个NIH和1个nsf资助的研究项目，这些项目涵盖了广泛的主题，所有这些项目都从TEM和免疫电子显微镜的分析中受益匪浅。该仪器的主要用户所涉及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中肌动蛋白细胞骨架的结构和功能（Bretscher博士），酿酒酵母中微管在染色体分离中的作用（Huffaker博士），劳斯肉瘤病毒的组装和包装（Vogt博士）。其他也受益于TEM分析的研究课题，但在不太一致的基础上，因此形成了一个次要的用户组，包括非洲爪蟾和酵母有丝分裂检查点对中期到后期转变的调节（陈博士），核层蛋白Ya的作用，它是受精卵中从减数分裂到有丝分裂的过渡所必需的（沃尔夫纳博士），秀丽隐杆线虫早期胚胎极性的建立（Kemphues博士），以及胆固醇在膜结构中的作用（费根森博士）。这个小组的成员将继续在他们的研究中使用TEM，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和免疫电子显微镜进行分析。

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.