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，而且还需要确定这些途径的空间和时间组织。成像核心的目的是为首席调查员在这方面的调查提供支助。一般而言，核心将提供咨询、培训和获得获取和分析光学显微镜下细胞结构所必需的设备。核心支持四大类显微技术：1)亚细胞的鉴定 利用常规光学显微镜和共聚焦显微镜观察分子的分布；2)使用共聚焦光学平面的组装或去卷积方案对免疫荧光标记的细胞进行三维重建；3)在活细胞中显示荧光融合蛋白；以及4)在固定细胞和活细胞中进行荧光共振能量转移(FRET)。后两种技术代表了Core中可用的最专门和最强大的技术，因为它允许在活细胞中实时确定多个信号分子的共同定位。FRET将被用于体内选择的激酶的测定和 小G蛋白的活性以及关键信号中间产物的共定位。

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.