The central goal of this program is to develop new and effective gene therapies for cardiovascular disease. The Biologic Imaging Core will play multiple diverse roles within this program. Visualization and localization of message, protein, or structural change resulting from gene expression is an essential step in evaluating the efficacy of successful gene transfer into cells and tissues. This identification varies from the low resolution studies of whole tissues, defining correction of pathological phenotype, to high resolution observations of successful sub-cellular passaging and presentation of protein. These studies will employ the full array of current light, and potentially, electron microscopic methods including: single and multi-color fluorescence microscopy, laser confocal microscopy, digital deconvolution, live cell imaging, transmission electron microscopy and computer-aided morphometric analyses. The Center for Biologic Imaging, in which this core service will be performed, is designed for the purpose of providing state of the art microscopic technologies to its users. It i equipped to perform a continuum of optical methods including all types of light and electron microscopy essential to this Program Project. Within the scope of this project at the light microscopic level these methods include: histology, immuno-histology, live cell and in situ hybridization methods. At the electron microscopic level we will provide fine structural and immuno-electron microscopic evaluation of specimens as a natural extension of the light microscopic analyses. Furthermore, our considerable experience in computerized image processing and morphometry will allow quantitative analyses of observed phenomena to corroborate qualitative changes in molecular expression, cell number or morphologic appearance. This core will be used by all projects, through the imaging tools used will vary from project to project. Preliminary data have shown the validity of these approaches and we expect a very significant increase in the use of optical techniques within the formal setting of this program.
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