High Pressure Freezer

高压冷冻机

• 批准号: 7387514
• 负责人: LUCILLE SHAPIRO
• 金额: $ 24.54万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7387514
• 关键词: Animal Model; Area; Arvin; B-Lymphocytes; Bacterial Chromosomes; Biological; Biological Preservation; Biology; Cell Polarity; Cells; Complex; Cytoskeleton; Development; Disease; Electron Microscopy; Electron Microscopy Facility; Freezing; Funding; Gold; Health; Human; Image; Institution; Light; Methods; Microtubules; Molecular; Nematoda; Neurons; Range; Regulation; Request for Proposals; Research; Resolution; Resource Sharing; Rodent; Sampling; San Francisco; Science; Sensory; Standards of Weights and Measures; Structure; Subcellular structure; System; Techniques; Tissues; Touch sensation; United States National Institutes of Health; Universities; Viral; Viral Cancer; Viral Pathogenesis; Yeasts; Zebrafish; base; human tissue; improved; in vivo; neural circuit; pressure; receptor; research study; response; sample fixation; synaptogenesis

DESCRIPTION (provided by applicant): This proposal requests funds to purchase a Leica EM PACT2-RTS high pressure freezer for rapid, cryo- fixation of biological samples. The high pressure freezer will be a shared resource, located in a well- established, multi-user, light and electron microscopy facility at Stanford University: the Cell Sciences Imaging Facility. This facility is accessible to the entire Stanford campus as well as surrounding institutions such as UC Santa Cruz and UC San Francisco. High pressure freezing has become the gold standard for fixation of biological electron microscopy samples. In the numerous studies were it has now been applied, this method has extended our understanding of the structural and molecular organization of cells and tissues. High pressure freezing provides much improved fixation, both in terms of quality and quantity, over all other fixation techniques and, when used in conjunction with a rapid transfer system, provides increased temporal resolution for correlative light to electron microscopy studies. For many studies and experimental systems high pressure cryo-fixation is not just desirable but necessary and essential for proper preservation of cellular fine structure and antigenicity. The high pressure freezer will support NIH funded projects from eight major users. These projects investigate a wide range of NIH supported topics, including, 1) mechano-electrical transduction and microtubule cytoskeleton organization in nematode touch receptor neurons (Goodman); 2) microtubule cytoskeleton organization and regulation (Stearns); 3) molecular basis of T and B lymphocyte recognition and immuno-synapse formation (Davis); 4,5) molecular mechanisms of viral replication, assembly and egress from infected cells (Arvin, Kirkegaard); 6) development and function of the neural circuitry in Zebra fish, rodents and humans (Smith); 7) development of cell polarity in yeast (Pringle); 8) 3D organization of the bacterial chromosome and replisome (Shapiro, McAdams). These studies investigate critical structural questions in a variety of model organisms and human tissues and cover areas of research with implications for diverse aspects of human health and disease, ranging from cancer and viral pathogenesis to understanding the molecular basis of immuno-responses and sensory biology. All require levels of sample preservation that approach the in vivo state for ultrastructural study of subcellular structures and molecular complexes. Each and every one of these projects demands a level of fixation that approaches the native, hydrated state which is possible only by application of high pressure freezing.

描述（由申请人提供）：该提案要求提供资金购买 Leica EM PACT2-RTS 高压冷冻机，用于快速冷冻固定生物样品。高压冷冻机将是共享资源，位于斯坦福大学完善的多用户光学和电子显微镜设施：细胞科学成像设施中。该设施可供整个斯坦福大学校园以及加州大学圣克鲁斯分校和加州大学旧金山分校等周边机构使用。 高压冷冻已成为生物电子显微镜样品固定的金标准。在现已应用的众多研究中，该方法扩展了我们对细胞和组织的结构和分子组织的理解。与所有其他固定技术相比，高压冷冻在质量和数量方面都提供了大大改善的固定，并且当与快速转移系统结合使用时，可以为相关光电子显微镜研究提供更高的时间分辨率。对于许多研究和实验系统来说，高压冷冻固定不仅是可取的，而且是细胞精细结构和抗原性正确保存所必需和必需的。 高压冷冻机将支持 NIH 资助的八个主要用户的项目。这些项目研究了 NIH 支持的广泛主题，包括：1）线虫触觉受体神经元中的机电转导和微管细胞骨架组织（Goodman）； 2）微管细胞骨架组织与调控（Stearns）； 3）T和B淋巴细胞识别和免疫突触形成的分子基础（Davis）； 4,5）病毒复制、组装和从受感染细胞中排出的分子机制（Arvin，Kirkegaard）； 6）斑马鱼、啮齿动物和人类神经回路的发育和功能（史密斯）； 7) 酵母细胞极性的发展（Pringle）； 8) 细菌染色体和复制体的 3D 组织（Shapiro，McAdams）。 这些研究调查了各种模式生物和人体组织中的关键结构问题，涵盖了对人类健康和疾病各个方面具有影响的研究领域，从癌症和病毒发病机制到了解免疫反应和感觉生物学的分子基础。所有这些都需要接近体内状态的样品保存水平，以进行亚细胞结构和分子复合物的超微结构研究。这些项目中的每一个都需要达到接近天然水合状态的固定水平，而这只有通过应用高压冷冻才能实现。