Core--Imaging and Histology

核心——影像与组织学

• 批准号: 7410008
• 负责人: C. William Davis
• 金额: $ 15.69万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7410008
• 关键词: Adult; Animal Model; Arts; Biological; Biology; Biomedical Research; Clinical; Clinical Trials; Communities; Confocal Microscopy; Consult; Consultations; Controlled Clinical Trials; Count; Cystic Fibrosis; Development; Discipline; Disease; Electron Microscopy; End Point; Freeze Etching; Freeze Fracturing; Freezing; Frozen Sections; Gene Delivery; Hemophilia A; Hepatocyte; Histology; Image; Imagery; Imaging technology; Knowledge; Life; Light; Liposomes; Methodology; Methods; Microscopy; Modality; Modems; Molecular; Nasal Epithelium; Outcome Study; Photons; Plastics; Protocols documentation; Range; Rate; Research Personnel; Research Project Grants; Resources; Safety; Services; Software Tools; Specimen; Techniques; Technology; Testing; Therapeutic Studies; Training; Translational Research; Transmission Electron Microscopy; Treatment Efficacy; Viral Vector; Waxes; Work; base; cellular transduction; cystic fibrosis airway epithelia; cystic fibrosis patients; design; digital; digital imaging; double-blind placebo controlled trial; epon; experience; gene therapy; improved; instrumentation; light microscopy; novel; novel therapeutics; programs; technique development; therapeutic transgene; tissue processing; transgene expression; vector

Gene therapy as a discipline cuts across the entire spectrum of biomedical research, and a successful program requires access to all of the technical resources available to modern biology. As one of these technologies, biological imaging, including histology, has made major contributions to our biomedical knowledge in recent years as the instrumentation and techniques available have become not only more sensitive, more diverse, and more robust, but also easier to use. We have developed within the past year the wholly new Michael Hooker Microscopy Facility which will allow the Imaging & Histology Core to offer to the UNC gene therapy research community a suite of state-of-the-art technologies and technical assistance to aid their work. UNC's gene therapy research focuses on an expansion of knowledge of molecular mechanisms involved with the delivery and permanent expression of the therapeutic transgenes, with the long range objective of providing novel therapeutic modalities for treating monogenetic diseases such as cystic fibrosis and hemophilia. From the experience of two controlled clinical trials, the UNC Program has established the following objectives, 1) translational research with defined clinical endpoints, that provide a basic understanding of efficient gene delivery to airway epithelia for CF and liver cells for hemophilia; 2) the development of high titer viral vectors that offer safe, efficient long-term transgene expression; and 3), the development of novel animal models to help us better understand rate limiting steps in target cell transduction. The first and last of these requires heavy utilization of imaging and histology which the Imaging & Histology Core will serve with specific aims pertinent to the provision of [i] digital based widefield and confocal microscopy and cryo-cooled CCD-based luminometry, [ii] electron microscopy (including freeze fracture/freeze etch), [iii] tissue processing and sectioning for frozen specimens, or embedding in wax or plastic as appropriate, for light microscopy, and [iv] tissue processing and ultrathin sectioning services for embedding in plastic as appropriate to transmission electron microscopy. The Core will also assist investigators with the development of techniques for the expression and visualization of probes within living specimens or of novel or improved histological methodologies, and Core staff will consult with gene therapy investigators on methods of image quantitation.

基因治疗作为一门学科跨越了生物医学研究的整个领域，一个成功的项目需要获得现代生物学可用的所有技术资源。作为这些技术之一，包括组织学在内的生物成像近年来为我们的生物医学知识做出了重大贡献，因为可用的仪器和技术不仅变得更加灵敏、更加多样化、更加稳健，而且更易于使用。我们在过去的一年里开发了全新的迈克尔·胡克显微镜设施，该设施将使成像和组织学核心能够向北卡罗来纳大学基因治疗研究界提供一套最先进的技术和技术援助，以帮助他们的工作。北卡罗来纳大学的基因治疗研究重点是扩展分子知识 涉及治疗性转基因的传递和永久表达的机制，其长期目标是为治疗囊性纤维化和血友病等单基因疾病提供新的治疗方式。根据两项对照临床试验的经验，北卡罗来纳大学项目确立了以下目标，1）具有明确临床终点的转化研究，为CF的气道上皮和血友病的肝细胞的有效基因传递提供基本了解； 2) 开发高效价病毒载体，提供安全、高效的长期转基因表达； 3）开发新型动物模型，帮助我们更好地了解靶细胞转导中的限速步骤。其中第一个和最后一个需要大量利用成像和组织学，成像和组织学核心将服务于与提供[i]基于数字的宽场相关的特定目标 和共焦显微镜和基于冷冻冷却 CCD 的发光测定法，[ii] 电子显微镜（包括冷冻断裂/冷冻蚀刻），[iii] 冷冻标本的组织处理和切片，或酌情包埋在蜡或塑料中，用于光学显微镜， [iv] 适合透射电子显微镜的组织处理和塑料包埋超薄切片服务。核心还将协助研究人员开发活体标本中探针的表达和可视化技术或新颖或改进的组织学方法，核心工作人员将就图像定量方法与基因治疗研究人员进行协商。