High-parametric flow cytometry cell sorter for human immunology studies.

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PROJECT SUMMARY Flow cytometry is the most commonly used and critical tool for analyzing immune cell populations in tissues, enabling approximately 18-20 parameter separation of populations and maintaining cell viability. By passing cells through a stream in which laser excitation is applied, individual populations of cells can be separated into either 96 well plates as single cells, enabling cloning and other applications, or can be enriched as populations in 4-6 separate populations simultaneously. This is the only cell separation technology universally adaptable to multiple tissues including isolation of rare populations from difficult sites such as peripheral tissues, necrotic tumors, and organs. Currently, this cutting-edge technology is not readily available to the Weill Cornell Medical College (WCMC) and medical center research community. Our current flow cytometry core is at maximum capacity usage for basic science applications and is booked weeks to months in advance, prohibiting alignment to the needs of clinical research investigators and basic and translational research directly on patient samples. Based on a prospective survey of WCMC scientists, we have identified over 12 NIH-funded major users, as well as a large number of minor users from multiple departments who have requested this instrumentation to advance their work. The proposed applications are diverse, leveraging the various scientific strengths of a number of WCMC labs. Some examples include Dr. Ari Melnick's work on understanding human B cell germinal centers, Dr. Sallie Permar’s understanding of pediatric infections and human vaccine responses and correlates of protective immunity, Dr. Laura Santambrogio’s ongoing studies on the human MHC peptidome to understand immune antigen presentation, and Dr. Brad Jones’ investigations into HIV pathogenesis and T cell immune responses. The leadership of WCMC is strongly supportive of the acquisition of this instrument, and will provide significant additional funds beyond those requested in the S10 application. In addition, dedicated space has been allocated within the Belfer Research Building and Human Immune Monitoring Core, where the instrument will be located and managed. Overall, we hypothesize that the establishment of a Flow Cytometry cell sorter instrument on the WCMC campus will be pivotal in driving new scientific insights across a number of projects and disciplines.