This Shared Instrument Grant application is for the purchase of a high-speed fluorescence-activated cell sorter, FACSAria II by Becton Dickinson. This instrument is peerless in its user friendly design and offers maximal flexibility to adjust to various experimental needs;thus it is an ideal choice for user-operated shared equipment. We have custom configured the FACSAria II to have four lasers at 405 nm, 488 nm, 561 nm, and 640 nm and twelve photomultiplier tubes for simultaneous analysis of ten fluorescent parameters. The custom FACSAria II also comes with a fully enclosed fluidic system and an advanced aerosol management option that meets the safety standard for processing samples contaminated with human pathogens. The impetus for this application is insufficient cell sorting capacity currently available, increased cell sorting demand, and the lack of appropriately configured devices for new research developments at the University of California, San Francisco (UCSF). The new instrument has a projected usage of over 2500 hours per year by fifteen defined investigators, demonstrating a real need for this instrument. Six of the investigators are new and early-career investigators, two are new to UCSF Parnassus campus, and three are new users of cell sorters due to new project development;together these new activities represent over 70% of the projected use of this instrument. These new activities cannot be fully accommodated by existing, already oversubscribed sorters at UCSF. Furthermore, clinical and human sample processing represent over 40% of the estimated usage of this machine. Lastly, in addition to the general shortage of the sorters, there is very limited capacity to sort cells based on red fluorescence due to the lack of green lasers on existing sorters. In response to these needs, we have configured the new sorter with improved aerosol containment for human sample processing and an additional yellow-green laser for sorting cells with red fluorescent tags. Thus, the new sorter is tailored to maximally meet new research demands at UCSF. Researchers directly benefiting from this instrument are from five departments in diverse fields ranging from molecular and cellular research in animal models to translational and clinical projects. This instrument will directly support research projects by 9 investigators with active NIH funding and 6 new early career investigators. The new sorter will be jointly managed by the Laboratory of Cell Analysis (LCA) and the Transplantation Research Laboratory (TRL). The LCA is a campus-wide core program that has over sixty years of experience in flow cytometry support. It will be responsible for technical and administrative support of the instrument. The TRL will house the instrument and provide backup onsite technical support. The new instrument will be integrated into the existing network of sorters managed by LCA to ensure expert maintenance of the instrument. Coordinated management of similar equipment will also ensure consistency in sorter recharge rate and access across the campus, which will likely benefit the UCSF research community at large.

Public Health Relevance

We are requesting a high-speed Sorter to be shared by more than 15 researchers at UCSF Parnassus campus. This instrument will support research in a wide range of research fields such as Transplantation, Diabetes, Asthma, Cancer, and Immunology.

National Institute of Health (NIH)
National Center for Research Resources (NCRR)
Biomedical Research Support Shared Instrumentation Grants (S10)
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Special Emphasis Panel (ZRG1-CB-J (30))
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Levy, Abraham
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University of California San Francisco
Schools of Medicine
San Francisco
United States
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Klementowicz, Joanna E; Mahne, Ashley E; Spence, Allyson et al. (2017) Cutting Edge: Origins, Recruitment, and Regulation of CD11c+ Cells in Inflamed Islets of Autoimmune Diabetes Mice. J Immunol 199:27-32
Putnam, A L; Safinia, N; Medvec, A et al. (2013) Clinical grade manufacturing of human alloantigen-reactive regulatory T cells for use in transplantation. Am J Transplant 13:3010-20