The cellular morphology core laboratory is constructed to met the increasing demands of cell biology and transgene detection in the area of gene therapy research. Routine morphology support in the form of frozen, paraffin, and glycolmethecrylate sectioning for light level microscopy (LM) as well as standard transmission electron microscopy (EM) will be provided as a comprehensive service to investigators participating in the CF core center grant. In addition, the core will emphasize state of the art methodologies for detecting transgene expression including in situ hybridization, immunocytochemistry, and histochemical detection of beta- galactosidase and alkaline phosphatase reporter genes. Within initial phases of pilot projects, these techniques will be provided as comprehensive services in order to evaluate feasibility, within a setting of minimal technical error. Following evaluation of feasibility to detect gene transfer in the various projects, an emphasis will be placed on transferring technologies to outside users. During this second phase of pilot projects, users will have full support for routine sectioning and full access to equipment and commonly used reagents for specialized techniques such as in situ hybridization, immunocytochemistry, and histochemistry. This decreased burden on the core to provide a comprehensive service for specialized techniques throughout the entire life of pilot projects will allow the core to also place substantial efforts on the development of new and innovative ways to detect gene expression. These developmental efforts will allow investigators to address new hypotheses about cell biology not currently possible. Specifically, the core plans to develop methods for techniques of insitu detection of mRNA at the EM level as well as dual fluorescent insitu hybridization at the LM level.
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