Image Analysis Core

对控制淋巴细胞分化和功能的信号转导过程的理解 不仅需要描绘精确的生化事件链，而且还需要确定 和时间组织的途径。Imaging Core的目的是为 在这方面进行调查的主要调查员。一般而言，核心将提供咨询、培训和 获得在光学显微镜下获取和分析细胞结构所必需的设备 水平核心支持四大类显微镜技术：1）亚细胞识别 使用常规的光和共聚焦显微镜的分子分布; 2）三维 使用共聚焦光学平面的组装重建免疫荧光标记的细胞，或 去卷积方案; 3）活细胞中荧光融合蛋白的可视化，和4）荧光融合蛋白的可视化。 共振能量转移（FRET）在固定和活细胞。最后两个代表了最专业的， 核心中可用的强大技术，因为它允许确定多个信令的共定位 分子在活细胞中的真实的时间。FRET将用于所选激酶的体内测定， 小G蛋白活性以及关键信号传导中间体的共定位。

The understanding of signal transduction processes governing lymphocyte differentiation and function require not only a delineation of the precise chain of biochemical events, but also determination of the spatial and temporal organization of the pathways. It is the purpose of the Imaging Core to provide support for the Principal Investigators in investigations in this regard. In general, the Core will provide advice, training and access to equipment essential for the acquisition and analysis of cell structure at the light microscopical level. The Core supports four broad categoriesof microscopic techniques: 1) identification of the subcellular distribution of molecules using conventional light and confocal microscopy; 2) three-dimensional reconstruction of immunofluorescent-labeled cells using assembly of confocal optical planes or deconvolution protocols; 3) visualization of fluorescent fusion proteins in living cells and 4) fluorescence resonance energy transfer (FRET) in fixed and live cells. The last two represent the most specialized and powerful technique available in the Core, as it allows the determination of co-localization of multiple signaling molecules in real time in living cells. FRET will be utilized for both the assay in vivo of selected kinase and small G protein activities as well as for the co-localization of critical signaling intermediates.