The Flow Cytometry and Cellular Imaging Core Facility (FCCICF) provides cellular analysis to investigatorswith peer-reviewed grants. The FCCICF has two sites, on the north campus and on the recently-openedsouth campus. It occupies 1900 sq. ft. and is directed by Dr. Michael Andreeff. The FCCICF develops andprovides techniques for single-cell analysis. Cell phenotyping, proliferation, signaling and apoptosis assayshave been established and modified for multiparametric analysis. Immunophenotypic analysis was combinedwith assays of intracellular proteins related to apoptosis (Bcl-2, Bcl-XL, BAG-1, p53, Rb, caspase activation,mitochondria! membrane potential and Fas), cell signaling (MARK), proliferation (Ki67, cyclins, BrDU, PCNA,DNA) and cell division history (CFSE). Quantitation of cellular antigens allows determination of the antibodybindingcapacity per cell. Rare events and progenitor/stem cell subpopulations can be detected and isolatedby three-laser excitation/eight-parameter fluorescence-activated cell sorting (FACS) for subsequent analysisby molecular cytogenetic and other molecular techniques including quantitation of intracellular PKCoc, Bax,Bcl-2, ERK, pERK, XIAP by laser scanning cytometry. FISH has been combined with apoptosis assays todiscriminate apoptosis in normal and malignant cells. The number, phenotype and proliferation of minimalresidual disease cells can be determined at levels of one malignant in 30,000 normal cells. Methods fordetection of transgene expression in cells are in place using p-galactosidase (p-gal), nerve growth factorreceptor (NGF-R), and green fluorescent protein (EGFP). Acquisition of 3 new FACS Aria cell sorters and afour-laser flow cytometer (B&D LSRII) has upgraded the facility to provide state-of-the-art isolation andanalysis. Laser confocal microscopy has been used extensively and was upgraded by acquisition of anOlympus FV-500 multi-user instrument and a DSU spinning disc confocal system. High-impact studies incancer prevention, growth factor signaling in breast cancer, apoptosis regulation in leukemias and multisteptumorigenesis all utilized confocal microscopy. The FCCICF now utilizes 17 major instrument systems. TheCore supports numerous R01, P01, R21, and SPORE grants. Since the previous review, the number ofusers has increased 145% (169 investigators with peer-reviewed grants from 19 different programs). 67% ofusers have peer-reviewed funding. During the previous funding period, 11,668 hours of service wasprovided. Use has tripled to greater than 7,000 hours estimated for 2007. Future plans are focused on thecontinued development and application of cutting-edge technologies in cell analysis.
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