ShEEP Request for Fluorescence-Activated Cell Sorter (FACS)

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The project is a ShEEP request for a Becton Dickinson (BD) FACSMelodyTM Cell Sorter. The BD FACSMelodyTM Cell Sorter is a high quality flow cytometry sorting instrument, with an automated instrument setup routine, that has the advantage of being easy to learn and use. The unit provides researchers a supportive and comprehensive workflow experience, and is derived from the cited and proven BD FACS Aria Cell Sorter. Typically the system is ready in less than 17 minutes- maximizing uptime. The small unit creates streamlined workflow with instrument settings, as well as gates and sort settings available and retrievable from saved experiments. The cell sorter system anticipates potential issues and identifies root causes with filter/mirror units that communicate with the software to confirm the optical configuration supports for experiments. The instrument will facilitate individual VA investigators with widely varying projects to isolate individual types of cells from normal and disease tissue and study their characteristics, leading to advances in the understanding of tissue inflammation, cancer immunology, and nervous system regeneration. The characteristics of the instrument make it ideal for our VA Research Service because this award will support the work of 5 major VA Merit-funded users (Drs. Hornyak, Atamas, Luzina, Davila, and Makar), and a minor user (Dr. Golding) who is a Career Development Award (CDA-2) recipient. Thomas J. Hornyak, M.D., Ph.D., is the Principal Investigator (PI) for this proposal. He is the Associate Chief Of Staff (ACOS) for Research and Development (R&D) at the VA Maryland Health Care System (VAMHCS), and PI on the BLR&D Merit Review project “Melanocyte Stem Cells in Regenerative Medicine”. He and his laboratory will use the sorter to isolate subsets of both mouse and human skin-derived stem cells in the viable state to evaluate their properties to promote neural regeneration. Sergei Atamas, M.D., Ph.D., and Irina Luzina, M.D., Ph.D., are individually funded VA Merit Review investigators with laboratories located in the Baltimore VA Medical Center (VAMC) who collaborate on the study of interleukin (IL)-33 and pulmonary fibrosis. They will sort tissue- derived macrophages and lymphocytes with the FACSMelodyTM. Eduardo Davila, Ph.D., is a VA Merit Review investigator whose research program is focused on tumor immunology. He will use the cell sorter for his VA Merit Review project to isolate tumor cells expressing varying levels of IRAK-4, postulated to mediate tumor resistance to therapy. Tapas Makar, Ph.D., is a VA Merit-funded investigator who is interested in determining the ability of brain-derived neurotrophic factor (BDNF) to modulate murine experimental autoimmune encephalitis, a disease model for autoimmune demyelinating disease in humans. He will sort immune and inflammatory cells from mouse spleen after experimental treatments to evaluate the immunosuppressive side effects of those treatments. Amit Golding, M.D., Ph.D. is a VAMHCS rheumatologist and a CDA awardee that treats Veterans in the Baltimore VAMC Arthritis Clinic. He submitted a VA BLR&D Merit Review application (Fall 2016 Cycle) subsequent to his CDA-2 project; and has a research program emphasizing autoimmune disease. Dr. Golding will sort B lymphocytes and T regulatory cells from serum samples from patients with autoimmune disease and normal controls for his studies.