This proposal requests funds to purchase a Cytomotion MoFlo high speed, high purity flow cytometric cell sorter to support the research of nine (9) Biomedical research faculty with a total of 11 NIH supported projects at North Carolina State University. The instrument will be housed in the Flow Cytometry Core Facility at NCSU. Some of the projects focus on small subsets of T lymphocytes that regulate resistance or immunodeficiency in the FIV-model for HIV-induced AIDS. Two projects examine bronchial epithelial cell populations and their functions. Another project includes utilizing knock-out mice to study the factors of cells involved in chronic inflammatory bowel disease; utilizing transfected mutated transcription factors to analyze expression of specific endo cell-cycle genes regulating susceptibility to retinoblastoma; and gene targeting and animal cloning technology. In all cases, MoFIo technology is required to separate cell subsets based on either surface markers or cell cycle to analyze specific proteins and/or cell function. The PIs currently have available a FACSCaliber for cell sorting, but its sorting speed (500 cells/sec) to too slow. The MoFIo sorts at a rate up to 70,000 cells/sec, and has the capability of simultaneous sorting of up to 4 cell subsets from a mixed population. There is a MoFIo at the University of North Carolina-Chapel Hill and two FACSVantage sortesr at Duke University, both 30 miles from NCSU. The difficulty in scheduling these instruments and long commuting time (minimum of 45 minutes one way at best) makes it impractical to use this technology on a routine basis. The University will provide support for the MoFIo operator and costs to place the instrument in operational order. Users fees established by the MoFlo oversight committee will cover subsequent service contracts. Client use of the MoFIo will be determined by the oversight committee under current guidelines for the Flow Cytometry Facility. The PIs on this proposal will be given priority; however, other faculty will be encouraged to use the technology as the need arises. ? ?
