The Core is actively involved in a variety of applications including the isolation of tissue and blood cell populations for animal transplantation experiments; isolation of cell sub-populations for analyses of functional cellular properties and gene transcription profiles, as well as high throughput screening of enhancer regions. Instrumentation: The Core is equipped with two Becton Dickinson FACSARIAs which can identify up to 13 fluorescent parameters and sort into tubes, 96 or 386 well plates, or slides; one Becton Dickinson LSRII which is used for direct high dimensional flow cytometry analysis applications (13 color), including high throughput analyses; and one four color BD FACSCalibur for more basic analyses. The Coulter CytoFLEX S was recently added to the Core's repertoire of instrumentation and should be a welcome addition for investigators as it is capable of thirteen color analysis and is equipped with a plate reader. The Flow Core maintains a laser scanning cytometer (LCS). Current applications (100%) are focused on zebra fish projects including drug screens and blood development. An autoMACS Pro Separator is used for high-speed magnetic cell isolation of multiple samples. The autoMACS Pro Separator is designed for cell isolation in a fully automated, walk-away fashion. Current issues: We have upgraded both sorters to have 13-color capability and have introduced index sorting and added an additional analyzer. The index-sorting feature enables the investigator to sort cells into single wells of plates, and easily identify them on plots, in statistics views, and in a detailed results file. These features allow the rapid isolation and characterization of single cells for use in single cell RNA-seq, Fluidigm and other genomic analyses. The Core is approved as a BSL2 laboratory and is currently sorting of samples or samples from virally infected mice. A concern is that the cell sorter instruments are aging (one sorter is fifteen years old and the second is twelve). Both instruments are covered by service contracts. These instruments are heavily used by many investigators in the Institute for isolation of unique cell populations from a wide variety of sources and will face increased down time with age. The Flow Cytometry Core is currently staffed by Stacie Anderson (Core Director, 100% effort, GMBB), and Martha Kirby (100% effort, GMBB). These investigators run the sorters, maintain the analyzers and other equipment and provide expert advice on design and execution of experiments.
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