The purpose of the Technical Support Core Facility is to provide the technological expertise to: quantitate cellular subsets in cell suspensions derived from reproductive tract tissues; determine the expression of cytokines in and their receptors on cells derived from reproductive tract tissues; and analyze the expression of cytokine mRNA and cell surface antigens in tissue sections from the reproductive tract. This Core Facility is composed of three units: Flow cytometry services; cytokine analysis services; and in situ analysis/image analysis services. This facility has been developed as a central service for Program Investigators to provide standardized technology to meet specific scientific needs. The functions of the three components of the facility can be summarize as follows. The Flow Cytometry Service will perform comparative multiparameter flow cytofluorometric analysis on single cell suspensions from reproductive tract tissues and peripheral blood cells. This facility will maintain a panel of monoclonal antibodies and, using titered amounts of these antibodies, stain and analyze samples on the cytometer and report data. In addition, this facility will prepare samples for sterile and non-sterile sorting and sort cells into required sub-populations. The Cytokine Analysis Service will provide a multi-tiered approach to the quantitation of cytokine protein and mRNA. Real Time PCR will be used to quantify mRNA levels for cytokines and cytokine receptors. Bioassays that specifically measure biologically active cytokines will be used for the detection of cytokines in tissue culture fluids. Finally, flow cytofluorometric analysis of cytokineexpressing cells will be provided. The Immunohistochemistry and In situ Analysis Service will provide the technology for the immunohistochemical and in situ analysis of cell surface antigen expression and the identification of cytokine-producing cells. With the recent success in confocal image analysis, expanded support for this technology will be provided.
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