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