CELLULAR AND MOLECULAR MORPHOLOGY CORE

细胞和分子形态核心

• 批准号: 8611913
• 负责人: Milton j Finegold
• 金额: $ 21.91万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-15 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8611913
• 关键词: 3-Dimensional; Address; Antibodies; Basic Science; Biological Assay; Cell Fraction; Cells; Cellular biology; Clinical Research; Collaborations; Collection; Communication; Complex; Computers; Confocal Microscopy; Consultations; Coupled; Data; Data Set; Development; Digestive System Disorders; Disease; Dissection; Educational workshop; Environment; Enzymes; Equipment; Exhibits; Experimental Models; Fluorescence Microscopy; Fluorescent Dyes; Fluorescent Probes; Gene Chips; Gene Expression; Genes; Grant; Histocytochemistry; Histology; Housing; Human Resources; Image; Image Analysis; Immunization; Immunoelectron Microscopy; Immunofluorescence Immunologic; Immunohistochemistry; Immunologic Techniques; In Situ; In Situ Hybridization; Individual; Infection; Injury; Instruction; Internet; Intestines; Label; Laboratories; Laboratory Research; Lasers; Life; Medical center; Methodology; Methods; MicroRNAs; Microscopic; Microscopy; Molecular; Molecular Biology; Molecular Genetics; Monoclonal Antibodies; Morphology; Nucleic Acid Probes; Nucleotides; Optics; Organoids; Phase; Physiology; Procedures; Process; Productivity; Proteins; Proteomics; Protocols documentation; Publications; RNA; RNA Interference; RNA Splicing; Research; Research Personnel; Resolution; Resources; Scanning; Services; Shotguns; Staining method; Stains; Standardization; Surveys; System; Systems Biology; Techniques; Technology; Testing; Texas; Training; Translations; Variant; animal tissue; base; biological research; detector; digital imaging; experience; functional genomics; genetic analysis; high throughput screening; human tissue; hybridoma production; improved; interest; laboratory facility; light microscopy; locked nucleic acid; meetings; member; molecular pathology; novel strategies; operation; programs; research clinical testing; response; screening; time use; tomography; tool; transmission process

PROJECT SUMMARY (See instructions): The Cellular and Molecular Morphology Core provides expertise and specialized laboratory facilities to Gl researchers and develops new tests for clinical and basic research. This Core has been highly productive and beneficial for members of the TMC Digestive Diseases Center, particularly to Pilot and Feasibility (P/F) awardees, whose experience and resources are necessarily limited. In the last 4 years, 22 of the 50 Full Members used this Core as did 16 of 79 Associate members and P/F Awardees. Major services include histology, immunohistochemistry, in-situ hybridization, enzyme histochemistry and immunofluorescent antibody studies, confocal and deconvolution microscopy, transmission, scanning, and immunoelectron microscopy, quantitative morphometric analysis, laser capture dissection for molecular genetic analyses, and digital images for internet communication and publication. The Core provides consultation and training in collection and processing of human and animal tissues, as well as technical advice to researchers interested in developing sophisticated procedures. A major benefit of this Core is the sharing of information gained from diverse projects in individual laboratories to enhance the productivity of all. This is accomplished by workshops and meetings of the user groups at which new data, methods and procedures are presented. Advances during the past 4 years include: upgrading of the Laser Capture Microscopy system for phase and fluorescence microscopy; adding 40 new antibodies, some with double staining to meet investigators' needs; and the introduction of Optical Projection Tomography for 3-dimensional imaging of intestinal development and injury of intestinal organoids. High throughput imaging-based Monoclonal Antibody Screening was added in response to DDC members requests. This will be an efficient way to generate quality antibodies to purified proteins and antibody panels following 'shotgun immunizations' of multiple proteins (or cell fractions) which are then used in subsequent reverse proteomics approaches. Short DIG-labeled locked nucleic acid (LNA) probes have been introduced by the In-Situ Core. Since they are much shorter than conventional riboprobes (only 20-22 nucleotides long), LNA probes are applicable to micro RNAs and also detect the cellular localization of specific splice variants of RNA. Plans include standardization of a protocol to detect multiple genes at a time using riboprobes labeled with different tags and to combine RNA in situ hybridization and immunohistochemistry. This will be tested With fluorescent probes to permit confocal microscopy for exquisite cellular localization.

项目摘要（参见说明）： 细胞和分子形态学核心为 GL 研究人员提供专业知识和专门的实验室设施，并为临床和基础研究开发新的测试。该核心对 TMC 消化疾病中心的成员来说非常高效且有益，特别是对经验和资源有限的试点和可行性 (P/F) 获奖者而言。在过去 4 年中，50 名正式会员中的 22 名使用了该核心，79 名准会员和 P/F 获奖者中的 16 名也使用了该核心。主要服务包括组织学、免疫组织化学、原位杂交、酶组织化学和免疫荧光抗体研究、共焦和反卷积显微镜、透射、扫描和免疫电子显微镜、定量形态分析、用于分子遗传分析的激光捕获解剖以及用于互联网通信和出版的数字图像。该核心提供人类和动物组织收集和处理方面的咨询和培训，并为有兴趣开发复杂程序的研究人员提供技术建议。该核心的一个主要好处是共享从各个实验室的不同项目中获得的信息，以提高所有人的生产力。这是通过用户组的研讨会和会议来完成的，在这些研讨会和会议上提出新的数据、方法和程序。过去 4 年的进步包括： 升级了用于相位和荧光显微镜的激光捕获显微镜系统；添加 40 种新抗体，其中一些具有双染色以满足研究人员的需求；以及引入光学投影断层扫描技术，对肠道发育和肠道类器官损伤进行三维成像。应 DDC 成员的要求，添加了基于高通量成像的单克隆抗体筛选。这将是一种有效的方法，可以在多种蛋白质（或细胞片段）的“鸟枪免疫”之后生成针对纯化蛋白质和抗体组的优质抗体，然后将其用于后续的反向蛋白质组学方法。 In-Situ Core 引入了短 DIG 标记的锁核酸 (LNA) 探针。由于 LNA 探针比传统的核糖核酸探针短得多（仅 20-22 个核苷酸长），因此适用于微小 RNA，还可以检测 RNA 特定剪接变体的细胞定位。计划包括标准化协议，以使用标有不同标签的核糖探针一次检测多个基因，并将 RNA 原位杂交和免疫组织化学结合起来。这将使用荧光探针进行测试，以允许共聚焦显微镜进行精细的细胞定位。