Cell and Tissue Imaging Core

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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. The goals of this program are to extend our understanding of the cellular pathology of Duchenne Muscular Dystrophy and to develop vector systems for dystrophin delivery using a variety of possible delivery systems, including AAV, Herpes and stem cells. It is expected that these vectors will be used to use these vectors to generate clinically useful therapeutic regimens. In each case there is a fundamental need to define the level of incorporation of the delivered dystrophin into muscle, and to assess the effectiveness of the therapy. This identification varies from the low resolution studies of whole tissues, defining correction of pathological phenotype, to high resolution observations of successful subcellular passaging and presentation of protein. The Center for Biologic Imaging, in which this core service will be performed, is designed with this function in mind. It is 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 include: histological, immuno-histological and possibly live cell technologies. 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 when needed. Furthermore, our considerable experience in computerized image processing and morphometry will allow quantitative analysis of observed _henomena to corroborate earlier, possibly quite subtle qualitative changes. This core will be used extensively by all projects, though 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