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，而且还需要确定途径的空间和时间组织。成像核心的目的是在这方面的研究中为主要研究者提供支持。一般而言，该中心将提供咨询、培训和获得在光学显微镜水平上获取和分析细胞结构所必需的设备。核心支持四大类显微镜技术：1）亚细胞识别 使用常规光和共聚焦显微镜观察分子的分布; 2）使用共聚焦光学平面或去卷积方案的组装对免疫荧光标记的细胞进行三维重建; 3）活细胞中荧光融合蛋白的可视化和4）固定和活细胞中的荧光共振能量转移（FRET）。最后两个代表了核心中最专业和最强大的技术，因为它允许在活细胞中真实的时间内确定多个信号分子的共定位。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.