Equipment for Visualization of Fragile Subcellular Detail by Electron Microscopy

通过电子显微镜观察脆弱亚细胞细节的设备

• 批准号: 7595603
• 负责人: MARCIA M MILLER
• 金额: $ 28.16万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-08 至 2010-04-07
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595603
• 关键词: Animal Model; Arts; Biological Preservation; Biomedical Research; Cities; Core Facility; Coupled; Cryoelectron Microscopy; Disease; Electron Microscope; Electron Microscopy; Ensure; Equipment; Event; Freeze Substitution; Freezing; Funding; Glutaral; Goals; Housing; Imagery; Intervention; Life; Malignant Neoplasms; Medicine; Methods; Minor; Organelles; Patients; Pharmaceutical Preparations; Preparation; Prevention; Research; Research Personnel; Sampling; Scanning; Specimen; System; Techniques; Translational Research; United States National Institutes of Health; Update; Vesicle; Work; cancer therapy; design; electron tomography; instrument; novel; pressure; public health relevance; quantum; sample fixation; tool; transmission process

DESCRIPTION (provided by applicant): Electron microscopy (EM) is an essential tool in present day biomedicine. Investigators often must provide EM evidence to support conclusions reached by other approaches. Recent advances in instrument design for high pressure freezing (HPF), one of the greatest advances in EM since the introduction of glutaraldehyde fixation, now allow HPF preservation of cellular ultrastructure to be more widely applied to a variety of cellular specimens in more labs. The superiority of preservation of ultrastructural detail in samples prepared by HPF freezing coupled with freeze substitution fixation (FSF) is readily evident, and this method is becoming the standard fixation method for EM. The goal of this proposal is to obtain funds for the purchase of a Leica EM PACT2 high pressure freezer, a Leica EM AFS freeze substitution instrument, and a Leica Microbiopsy Transfer System. This equipment will be housed in the EM Core Facility at City of Hope (COH) and will primarily be used by six Major Users and four Minor Users, many of whom have R01 or other NIH funding. Many of the Major Users study cancer and are engaged in translational research focused on moving therapies from animal models to patients. They recognize the importance of optimal preservation of ultrastructure and have an increasing need for ultrastructural localization in their work. Some of these projects require immunolabeling for ultrastructural localization of molecules that could be lost or redistributed during conventional specimen preparation. Others need to distinguish endosomal and autophagic vesicles, organelles that are particularly sensitive to distortions during conventional sample preparation. The COH EM Core Facility was renovated in 2003, and was updated with new FEI Tecnai transmission and FEI Quanta ESEM scanning electron microscopes. We currently have a Leica cryo-ultramicrotome, and components for electron tomography and cryo-electron microscopy are being installed. The Leica EM PACT2 high pressure freezer, freeze substitution instrument, and microbiopsy transfer system will ensure that COH investigators have access to the most current fixation techniques for EM, which will greatly benefit their research on the prevention and treatment of cancer. Public Health Relevance: Contemporary medicine needs drugs that are rationally designed for specific interventions in cellular activities gone awry. Increasingly, biomedical research focuses on sub-cellular and macromolecular events to understand the details of cellular activities. The requested state-of-the- art Leica EM PACT2 high pressure freezer, Leica EM AFS freeze substitution instrument and Leica Microbiopsy Transfer System will aid NIH-funded investigators at City of Hope in developing novel therapies for cancer and other life-threatening diseases.

描述（由申请人提供）：电子显微镜（EM）是当今生物医学中必不可少的工具。调查人员通常必须提供EM证据，以支持其他方法得出的结论。自引入戊二醛固定以来，EM的最大进展之一，仪器设计的最新进展现在使HPF保留HPF的细胞超微结构可以更广泛地应用于更多实验室中的各种细胞标本。通过HPF冻结和冻结固定（FSF）制备的样品中的超微结构细节保存的优势很容易显而易见，并且该方法已成为EM的标准固定方法。该提案的目的是获得购买Leica EM PACT2高压冰柜，Leica EM AFS Free替代仪器和Leica Microbiopsy转移系统的资金。该设备将安置在Hope City（COH）的EM核心设施中，主要将由六个主要用户和四个未成年人使用，其中许多用户拥有R01或其他NIH资金。许多主要用户正在研究癌症，并从事转化研究，重点是将疗法从动物模型转移到患者。他们认识到最佳保存超微结构的重要性，并且在工作中对超微结构定位的需求越来越多。其中一些项目需要进行免疫标记，以进行分子的超微结构定位，这些分子在常规标本制备过程中可能会丢失或重新分布。其他则需要区分内体和自噬囊泡，这些细胞器对常规样品制备过程中对扭曲特别敏感。 COH EM核心设施于2003年进行了翻新，并通过新的FEI Tecnai传输和Fei Quanta Esem扫描电子显微镜进行了更新。目前，我们有一个leica冷冻粉状球体，并且正在安装电子断层扫描和冷冻电子显微镜的组件。 Leica EM PACT2高压冰柜，冷冻替代工具和微生物转移系统将确保COH研究人员能够使用EM的最新固定技术，这将极大地有助于他们对预防和治疗癌症的研究。公共卫生相关性：当代医学需要针对细胞活动的特定干预措施合理设计的药物。生物医学研究越来越重点关注亚细胞和大分子事件，以了解细胞活性的细节。所需的最先进的Leica EM PACT2高压冰柜，Leica EM AFS冻结替代工具和Leica微生物转移系统将有助于Hope的NIH资助的研究人员，以开发新的癌症和其他威胁生命的疾病的新疗法。