The central theme of this proposal is to dissect the molecular mechanisms leading to tissue injury, and organ dysfunction in trauma. Visualization and localization of message, protein, or structural changes leading to, or resulting from each step in this dynamic process is essential in understanding the molecular mechanisms of trauma. The different projects within this proposal focus on defined tissue systems including gut, liver and lung within models of shock and or trauma. In each case a wide range of microscopic methods will be employed and are considered essential in gaining an understanding the pathology of HS at the tissue and cellular level, thus a central cell and tissue imaging core is defined as an integral component of this proposal. The Core will be housed in the Center for Biologic Imaging (CBl) of the University of Pittsburgh Medical Center. This Center is equipped to perform a continuum of optical methods including all types of microscopy essential to this Program Project. Within the scope of this Program light microscopic techniques include: histological, immuno-histological, laser confocal, 2 photon, evanescent wave, live cell and whole dssue/animal imaging technologies. Our considerable experience in computerized image processing and morphometry will allow quantitative analysis of observed phenomena to corroborate subtle qualitative changes, and this a major function of the Core in this Program. At the electron microscopic level thin section electron microscopy and immuno-electron microscopic evaluation of specimens as a natural extension of the light microscopic analyses will be employed when needed. During the previous grant cycle the CBl collaborated extensively with all of project leaders, as is described in the preliminary data section and we expect a continued expansion in the use of optical techniques. Furthermore we have developed and built multiple new instruments to facilitate these interactions at all levels from the single cell to the whole animal
(See Instructions): Optical imaging is an essential core function within the program providing quantitative image based data for subsequent analysis using computer aided tools also available within the core
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