Core--Imaging /Histology

该核心的主要目标是为该项目组的研究者提供完整的专业知识、培训、试剂和设备，以完成固定和活细胞和组织中组织学、分子和mRNA靶点的成像。该核心支持PPG的五个研究项目，提供与光学检测，可视化和定量显微镜相关的服务。它提供了访问医学显微镜设施的约翰霍普金斯大学学院的完整资源的经验使用（其中包括一个优秀的电子显微镜设施）和医学部的多光子激光扫描显微镜设施。罗默博士和图德博士已经召集了人员和实验室设施，以满足广泛的实验成像需求。 核心人员有 在组织制备、细胞培养、差异去污剂提取和细胞骨架保存、细胞成分特异性染色、显微镜和数字图像分析等领域拥有丰富的专业经验。核心小组的技术人员为这一方案编制小组提供必要的支助。核心设施的特点是可以使用多个显微镜，这些显微镜利用Windows PC和Macintosh工作站，用于图像分析和可视化的广泛软件，以及用于制作复印就绪图像和硬拷贝数字的软件和硬件。该核心提供的具体能力包括：实验设计咨询、免疫标记和荧光蛋白嵌合体表达试剂、标本制备、活检处理、原位杂交、组织中放射性标记物质的放射自显影研究、光、荧光、多光子或电子显微镜免疫细胞化学标记 检查、透射光和旋转盘共聚焦实现二维和三维图像处理、分析算法和定量图像分析、三维重建和立体可视化以及涉及数字成像的图形服务。这个核心将准备和分析标本从广泛的研究工作。重要的是，在准备本提案的初步数据期间，该成像和组织学核心机构已证明其灵活性、专业知识和资源对于本PPG中的所有项目都是可用的和必不可少的。

The main objective of this Core is to provide the investigators in this program project group with a complete range of expertise, training, reagents, and equipment to accomplish the imaging of histologic, molecular, and mRNA targets in fixed and living cells and tissues. This Core supports the PPG's five research projects with services related to optical detection, visualization, and quantitative microscopy. It offers access to experienced use of the complete resources of the Johns Hopkins University School of Medicine Microscope Facility (which includes an excellent electron microscope facility) and the Department of Medicine's Multiphoton Laser Scanning Microscope Facility. Drs. Romer and Tuder have assembled the personnel and laboratory facilities to satisfy a wide range of experimental imaging needs. The Core personnel have professional experience spanning the fields of tissue preparation, cell culture, differential detergent extraction and cytoskeletal preservation, cell component-specific staining, microscopy and digital image analysis. The technical staff of the Core provide essential support to this PPG. The core facility features access to multiple microscopes that utilize both Windows PCs and Macintosh work stations, extensive software for image analysis and visualization, and software and hardware for producing manuscript-ready images and figures in hard copy. Specific competencies provided by this Core include: consultation on experimental design, reagents for immunolabeling and fluorescence protein chimera expression, specimen preparation, biopsy processing, in situ hybridization, autoradiographic studies of radio-labeled material in tissues, immunocytochemical labeling for light, epiflurorescence, multiphoton, or electron microscopic examination, transmitted light and spinning disc confocal implementation of two- and three-dimensional image processing, analysis algorithms and quantitative image analyses, three-dimensional reconstruction and stereoscopic visualization, and graphics services involving digital imaging. This core will prepare and analyze specimens from a broad range of research endeavors. Importantly, during the preparation of preliminary data for this proposal, this Imaging and Histology Core Facility has demonstrated that its flexibility, expertise, and resources are available and essential to all of projects in this PPG.