A HIGH PRESSURE FREEZER FOR ELECTRON MICROSCOPY USES

描述(由申请人提供)：本拨款申请的目的是 获得资金购买和安装一台高压冷冻机。 该装置将用于制备保存完好的断层扫描标本。 申请。高压冷冻提供最好的样品准备 一种保存细胞和组织超微结构的方法， 高分辨率现场资料。请求的设备是Bal-Tec 11 PM 010 高压冷冻机(HFP)和样品制备附件。 我们在结构生物学方面的研究项目涉及三维 用厚切片和中间体重建细胞和组织 电压电子显微镜(IVEM)。我们协调和整合我们的结构 研究数据来自分子显微镜，X射线晶体学， 生化、生物物理和神经生理学技术。我们的目标是 准确重建细胞和细胞器的结构 制备、显微镜和计算方法。目前，最近的HFP 公寓位于旧金山湾区或亚利桑那州图森市。HFP 将被安置在国家显微和成像研究中心 加州大学圣地亚哥分校。五大计划需要 HFP的用途包括： (1)缝隙连接膜通道的三维细胞组织 (2)结节细胞成分的结构研究 Ranvier(Ellisman，P.1)；(3)大分子缔合的变化 突触抑制时的小脑浦肯野神经元树突(Tsien 和Lev-Ram，P.1？S)；(4)无包膜病毒入侵机制(Johnson和 施尼曼，P.I.，S)；(5)三维结构和功能 电子断层扫描显示线粒体(Frey，P.1)。此外，还有四个 来自NCMIR研究资源的协作项目将立即 简要描述了使用HFP的好处。这九项研究 项目涉及超过25名调查人员，他们将在很大程度上依赖于 HPF。在所有这些研究项目中，主要目标是了解 有组织细胞组件的三维结构及其相关 这种形态与它们的功能有关。

DESCRIPTION (provided by applicant): The purpose of this grant request is to obtain funds to purchase and install a High Pressure Freezer (HPF) machine. This unit will be used to prepare well-preserved specimens for tomography applications. High pressure freezing delivers the best specimen preparation method for preserving cell and tissue ultrastructure and therefore, obtaining high resolution in situ data. The requested equipment is a Bal-Tec 11PM 010 High Pressure Freezing Machine (HFP) and accessories for sample preparation. Our research programs in structural biology involve the three-dimensional reconstruction of cells and tissues using thick sections and intermediate voltage electron microscopy (IVEM). We coordinate and integrate our structural studies with data obtained from molecular microscopy, X-ray crystallography, biochemical, biophysical and neurophysiological techniques. Our goal is to accurately reconstruct the structure of cells and organelles using the best preparative, microscopy and computational methods. At present, the closest HFP units are located in the San Francisco Bay Area or the Tucson, Arizona. The HFP will be housed in the National Center for Microscopy and Imaging Research at the University of California, San Diego. The five major programs requiring the use of the HFP are: (1) Three-dimensional cellular organization of gap junction membrane channels (Sosinsky, P.!.); (2) Structural studies of cellular components in the Node of Ranvier (Ellisman, P.1.); (3) Changes in macromolecular associations in cerebellar Purkinje neuron dendritic spines during synaptic-depression (Tsien and Lev-Ram, P.1?s); (4) Mechanisms of non-enveloped virus entry (Johnson and Schneemann, P.I.?s); and (5) Three-dimensional structure and function of mitochondria as viewed by electron tomography (Frey, P.1.). In addition, four collaborative projects from the NCMIR Research Resource that would immediately benefit from the usage of the HFP are described briefly. These nine research projects involve over 25 investigators that would depend heavily on the use of the HPF. In all of these research projects, the primary goal is to understand the three-dimensional structure of organized cellular components and relate this morphology to their functionality.