FLOW CYTOMETRY CORE

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Flow Cytometry Core will provide expertise in hematopoietic stem cell separations, microvesicle sorting, and cell phenotyping analysis. The Core will be headed by Mr. Mark Dooner. Mr. Dooner is an experienced flow cytometrist and will devote 80% time to this Core. A To Be Named Technician will be able to provide backup and cross coverage for the Core. We have decided to have an accomplished full time technician solely dedicated to this COBRE, since this is justified by the complexity and extent of the proposed work. This Core features an InFlux high speed cell sorter from BD Biosciences. This sorter is capable of carrying out stem cell separations, single cell deposition and is housed within a biosafety cabinet. This Core is housed in a dedicated room. The facility is equipped with a new LSRII (BD instruments) with FACS Diva software. Investigators will work with the core and prepare their cell populations for sorting or analysis. The Core will routinely separate Lineage negative Rhodamine lo Hoechst lo (LRH), LT-HSC, ST-HSC, CLP, CMP, LSK, SLAM, and Lineage negative Sca+ stem/progenitor populations. The core will also sort and characterize microvesicles and analyze populations for lineage profiles. The core will serve all projects in this COBRE carrying out both separations and analysis work. The Core also remains dedicated to the development of new separation techniques and the enhancement of known stem cell and microvesicle populations.