High parameter fluorescence activated cell sorter - Project Summary/Abstract The Versiti Blood Research Institute (VBRI) is an academic hematology research center located in Milwaukee, WI. The VBRI Flow Cytometry Core serves the needs of scientists at the VBRI, the Medical College of Wisconsin (MCW), and the Children’s Research Institute (CRI), three research-heavy institutions assembled on the Milwaukee Regional Medical Campus (MRMC). Currently, a heavily oversubscribed Cytek Aurora in the CRI is the only high parameter cell sorter on the MRMC. The VBRI’s 15-year old FACS Aria can no longer support the sophisticated sorting needs of many MRMC users and will need to be replaced in anticipation of BD’s discontinuation of servicing the product line. To address these deficiencies, we are requesting funds to purchase a Becton Dickinson Symphony S6 SE, a 5-laser, 49-detector cell sorter. The Symphony S6 matches the Symphony A5 SE analyzer purchased by VBRI in 2022, allowing for seamless transfer of multi-color panels from analysis to sorting. The research projects of our nine Major and eight Minor NIH-funded users require high parameter cell sorting to accomplish their goals. Using the Symphony A5 SE analyzer, these investigators have acquired valuable expertise in spectral flow cytometry, and the acquisition of a Symphony S6 will enable them to overcome critical roadblocks in their research. VBRI research will broadly benefit from this new instrument. In immunology, development of a B cell therapeutic for multiple sclerosis, the delineation of B cell involvement in heparin-induced thrombocytopenia as well as clarifying the mechanism of thrombotic complications in COVID- 19 all require access to high parameter sorting, using panels already established on the Symphony A5 SE. Major Users focused on hematologic malignancies require rapid access to a multi-parameter cell sorter to isolate rare cell populations from primary leukemia samples that can arrive any time. A signature program studying the role of the glycome in hematopoiesis is critically dependent on the Symphony S6’s increased number of channels. In thrombosis and hemostasis, studies on cellular integrins, kindlin signaling in neutrophils and platelets, and the pathophysiology of fetal/neonatal alloimmune thrombocytopenia will all heavily use the Symphony S6. Moreover, the Symphony S6 will overcome critical bottlenecks for MCW researchers studying the role of exosomes in ovarian cancer, chronic pain in Fabry and sickle cell disease, gamma herpesvirus associated B-cell lymphomas, immune response to radiation exposure and type I diabetes. FACS is an integral part of the workflow of most VBRI faculty and many researchers at MCW and CRI. Currently available high parameter (>20) sorting capacity is extremely limited, slowing progress on NIH-funded research. Even lower parameter sorting capacity is becoming limited on the MRMC as BD FACS Aria instruments are approaching end of lifespan. The Symphony S6 will provide essential high parameter sorting capacity for several NIH-funded projects, and dramatically accelerate progress on others. Ease of panel transfer, economic use of precious samples, and complementarity to existing instrumentation will expedite research progress to ascertain a high return on investment.
