The SIV Biosafety Level 3 (BSL3) / Flow Cytometry Core Laboratory will be directed by Dr. M. Christine Zink. The BSL3 technical staff will be responsible for processing all samples from the infected and uninfected control macaques throughout the experiments described in this Program and will provide samples and reagents to all projects in the program and perform measurement of markers on cells by flow cytometry (FACS). This core laboratory will fulfill the following Specific Aims: 1. To provide a safe work environment to perform work with infectious materials (ALL) 2. To provide support in the preparation and quantitation of virus stocks (ALL) 3. To receive, process, aliquot and log into appropriate databases all samples from macaques (ALL) 4. To isolate PBMCs, pDCs and mononuclear cells from blood (and lymph nodes, spleen and brain at necropsy) from animals for quantitation and longitudinal characterization of cells (ALL) 5. To isolate macrophages/microglia, astrocytes and lymphocytes from the brains of all infected and uninfected macaques for analysis. (Project 2 and 3) 6. To aliquot and archive plasma and CSF samples for viral load and other assays. (ALL) 7. Organizing, monitoring, and maintaining a physical inventory of specimens derived from all experiments {in vivo and in vitro), along with associated documentation and analytical reports (ALL) 8. To prepare samples for Real Time PCR (extraction of RNA or DNA). (ALL) 9. To establish primary cell cultures, and maintain cell lines. (ALL) 10. To perform, FACS experiments, analyze data and submit data to the database (ALL) 11. To property dispose of all the waste generated within the BL3 laboratory (ALL) 12. Maintain all equipment central to the function of the core (ALL)
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