Abstract: This proposal requests funds of $250K with remaining funds from our institution to purchase a high-parameter, state-of-the-art flow cytometer, BD FACSymphony A5 Cell Analyzer. This flow cytometer offers novel and cutting-edge technology aimed at simultaneous detection of up to 30 parameters for the study of many inflammatory diseases that include autoimmune, cardiovascular and neurodegenerative diseases as well as obesity, aging and cancer that are important areas of research in the COBRE on Dietary Supplements and Inflammation (DSI) at the University of South Carolina (UofSC). The ability to distinguish the multiple cell types based on biomarkers, receptors and functional characteristics as well as the ability to integrate the panels into a single composite that enables the diagnosis and prognosis of disease following attempts towards prevention and treatment with plant products will be most readily performed with this high-resolution and high-throughput instrument. This instrument will be housed in the Flow Cytometry and Imaging Core of the COBRE at the University of South Carolina, School of Medicine. This core will serve the needs of the Target Faculty and Pilot Project faculty as well as their graduate and undergraduate students, postdoctoral fellows and technical staff. The existing flow cytometers such as the FC500 are antiquated and offer a maximum of 5-color capability, thereby limiting the cell subpopulations that can be identified and have no data integration capability. In addition, these flow cytometers are not supported by the vendor as they are obsolete with parts not available. Further, FACSymphony has 5 lasers, 30 parameters and 28 color detectors offering high-performance detection capability with commercial, low cost, readily-available fluorochrome conjugated antibodies, high- speed processors, optimized photomultiplier tubes for each detector as well as updated software for analysis and visualization. Importantly, the instrument can be upgraded to 10 lasers, 50 parameters with 48 colors in the future. This bench-top instrument allows for rapid analysis of low volume of sample, can detect upto 28 colors, overcomes issues associated with fluorescence compensation and is user-friendly. As detailed in the proposal, the biomedical research environment in the COBRE on DSI at the UofSC is on a trajectory for expansion with no multi-parameter flow cytometer. The absence of this capability has adversely affected the research programs of the investigators participating in this COBRE as well as the recruitment of promising scientists to the UofSC. Acquisition of the instrument and housing it in an established core facility will allow access to it by a large number of investigators and will significantly enhance the research quality and training. Dr. Narendra Singh, a Research Professor and the Flow Cytometry, Microscopy and Imaging Core Leader has >20 years experience in running flow cytometry will oversee the operation of this system.

Multi-parameter flow cytometry is a relatively novel next-generation technology platform that can significantly advance our studies on inflammation that encompasses autoimmune, neurodegenerative and cardiovascular diseases as well as aging and cancer. Cells involved in disease progression express a complex array of components that indicate physiologic status, phenotypes, receptor expression and functional capabilities. The ability to distinguish the various types of cells while keeping the costs of reagents low is most readily performed with high resolution, multi-channel, high-throughput instrumentation such as the requested BD FACSymphony.