CORE--ADVANCED IMAGING SUPPORT

描述：（申请摘要）成像核心的主要目标是促进定量和定性成像技术在细胞和分子水平上分析蛋白质相互作用的使用。提供了广泛的成像技术，适用于研究细胞、组织、整个器官中的膜和细胞骨架蛋白。此外，转基因技术的应用为在全动物综合水平上研究表型变化提供了新的机会。新技术现在可用于特定目标蛋白的更高分辨率定位，以及允许动态变化可以识别的重要显微镜。石蜡切片光学显微镜，仍然是检查结构表型变化的金标准，已经扩展到包括使用免疫组织化学定位特定蛋白质和使用原位杂交和原位反转录定位特定mrna。共聚焦显微镜可应用于组织，其中可以使用离子敏感的荧光染料在组织水平上评估对细胞内离子浓度变化的分析，从而扩大在细胞水平上应用的视频成像技术的能力。利用免疫金和原位杂交技术提高了电镜免疫细胞化学技术的定位精度。数字图像分析系统的可用性增强了这些技术的价值，该系统允许定量形态测量和三维重建和可视化。这些技术在细胞病理学中非常有用，但对于没有受过培训或经验的调查人员和研究实习生来说，不容易获得。此外，选择适当的技术或技术组合需要在解释方面的经验和对方法局限性的了解。成像核心将在光、免疫荧光、共聚焦和电子显微镜的利用方面提供专业知识和协助，特别强调转基因模型的表型和病理分析、免疫细胞化学、生命显微镜和图像分析。该设施将提供这些专门技术和应用这些技术所需的设备，并将成为方案项目成员之间合作的协调中心。具体来说，该中心将为成像技术的有效应用提供必要的设施、服务和教育，以满足该项目各成员的个人研究目标。

Description: (Abstract of the application) The major goal of the imaging core is to facilitate the use of quantitative and qualitative imaging techniques in the analysis of protein interactions at the cellular and molecular level. A wide range of imaging techniques applicable to the study of membrane and cytoskeletal proteins in cells, tissues, whole organs are provided. In addition, the utilization of transgenic technology offers new opportunities to study the phenotypic changes at the integrated whole animal level. New technologies are now available for greater resolution of localization of specific target proteins as well as permitting vital microscopy where dynamic changes can be identified. Paraffin section light microscopy, still the gold standard for the examination of structural phenotypic changes, has been expanded to include localization of specific proteins using immunohistochemistry and of specific mRNAs using in situ hybridization and in situ reverse transcription. Confocal microscopy can be applied to tissues where analyses of changes in intracellular ion concentration can be assessed at the tissue level using ion sensitive fluorochromes resulting in an expansion of the capabilities of video imaging techniques which are applied at the cellular level. The utilization of electron microscopic immunocytochemical techniques has been improved for greater accuracy in localization using immunogold and in situ hybridization techniques. The value of these techniques is enhanced by the availability of digital image analysis systems that permit quantitative morphometric measurements and three-dimensional reconstruction and visualization. These techniques are extremely useful in cellular pathobiology but are not easily accessible to investigators and research trainees without previous training or experience. In addition the selection of the proper technique or combination of techniques requires experience in interpretation and knowledge of the limitations of the method. The imaging core will provide expertise and assistance in the utilization of light, immunofluorescence, confocal, and electron microscopy with particular emphasis on phenotypic and pathologic analysis of transgenic models, immunocytochemistry, vital microscopy and image analysis. The facility will give access to these specialized techniques and to the equipment necessary to apply them and will form a focal point for collaboration between members of the program project. Specifically, the core will provide the facilities, service and education necessary for the efficient application of imaging techniques tailored to the individual research objectives of the various members of the program project.