Project Summary Flow cytometry has been the gold-standard in biotechnology for interrogating cellular phenotypes for the past 60 years, and is limited only by the number of cellular parameters that can be simultaneously detected in samples. The capacity for multi-parametric flow cytometry using spectral-based systems has developed rapidly with the advancements in lasers and optics capable of more effectively exploiting the visible and invisible light spectrums. The requested BD FACSymphony A3 Cell Analyzer, with optional FSC PMT, is a state-of-the art five-laser (UV- 355nm, Violet-405nm, Blue-488nm, Yellow-561nm, Red-637nm) system capable of simultaneously acquiring data on as many as thirty cellular parameters and resolving particles as small as 200nm. Added advantage of this system is that it leverages the ease-of-use of classic BD instruments, which all Users listed in this proposal are experienced with, to create an advanced system with a simple user interface. Currently, the only system available at the University of South Carolina School of Medicine for performing routine cell analysis is a 12-year- old BD FACSAria II Cell Sorter. This proposal includes 8 Major Users with 16 NIH- and 2 VA-funded research grants. Our Users perform cutting-edge fundamental and translational research on a variety of human diseases including inflammatory bowel disease, lupus, atherosclerosis, Gulf War Illness, cardiomyopathy, cancer, and defects in hematopoiesis. The research programs of our Major Users will be benefitted by acquisition of this instrument for the following reasons: (1) increased ability to acquire more data per sample by inclusion of more lasers, (2) enhanced data quality by reducing spectral overlap through the integration of more lasers, (3) enhanced sensitivity to detect rare cell populations through integration of stronger lasers, (4) the ability to develop novel assays through inclusion of a yellow-561nm laser, (5) increased ability to discriminate cell populations by integration of stronger lasers and the ability to use stronger emitting UV and violet dyes, (6) the ability to develop novel assays for the enumeration of small particles (e.g. bacteria, exosomes, endosomes, mitochondria) by inclusion of special FSC PMT, (7) flexibility in customization of mirror/filter configurations for User-specific applications, and (8) the ability to use classic UV dyes (DAPI and Hoescht stains) in flow-based assays through inclusion of UV laser. Acquisition of this instrument advances the broad and long-term priorities of UofSC to become a leader in biomedical research in the state of South Carolina. First, it provides UofSC investigators an advanced platform for studying cellular responses. Second, it enhances the competitiveness of UofSC investigators for grant dollars by enhancing the complexity of questions that can be addressed experimentally, by providing a system capable of revealing unanticipated biological phenomenon that will allow investigato...