FLOW CYTOMETRY RESOURSE UPGRADE

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The FACS IV dual fluorescence/fifth parameter/computer upgrade requested here will enable the Flow Cytometry Laboratory at The University of South Alabama to perform the following functions not now available with the present flow cytometry ste-up: 1) simultaneous DNA and cell surface marker studies on viable cells, 2) two-color analyses using Texas Red as an alternative to phycoerythrin, 3) three-color analyses using Texas Red or allophycocyanin as the third fluorochrome together with FITC and phycoerythrin, 4) list mode data storage, making possible the re- analysis of samples, at a future date, using different gating parameters; also, making possible the simultaneous acquisition and storage of data from five parameters, 5) bivariate chromosome analysis using the Hoechst stains together with chromomycin A3, 6) contour plot, 3-D isometric, and multiple histogram data displays, 7) time of flight, and 8) single cell deposition for cloning and hybridoma screening. This upgraded FACS IV/computer resource will be instrumental in providing new capabilities for a variety of analytical and preparative cell techniques in the following NIH funded projects: 1. The characterization of oncofetal antigens, and monoclonal antibodies to these antigens, expressed on human and rodent tumors. 2. Analysis of a 44 kD oncofetal antigen on radiation induced leukemias and thymomas in mice. 3. Analysis and purification of parietal cells from the rat stomach. 4. Cell surface marker analysis of monoclonal populations of T helper cells, T suppressor cells and T contrasuppressor cells. 5. Phenotypic characterization and purification of T cell clones specifically reactive with Herpes simplex induced antigens. 6. Establishment of interferon- resistant cell lines and analysis of cellular receptors for interferon in the study of host defense against Rickettsiae. 7. Effect of alloantigenic challenge on lymphocyte sub-set composition in pregnancy and AIDS. 8. Isolation of insulin secreting beta cells from the pancreas. 9. Investigation of interferon-rickettsiae interactions using Rhodamine 123.