FLOW CYTOMETRY SHARED RESOURCE (FCSR): PROJECT SUMMARY The goal of the Flow Cytometry Shared Resource (FCSR) is to provide Wake Forest Baptist Comprehensive Cancer Center (WFBCCC) members with ready access to the necessary expertise and state -of-the-art flow cytometry instrumentation that can be applied to a wide range of cancer biology research. Flow cytometry is a critical technology that provides rapid single-cell analysis and cell sorting through the simultaneous detection of light scatter properties and the presence of multiple cellular molecules or metabolites in or on an individual cell. In addition, the FCSR seeks to foster the development of protocols that can utilize the continually increasing number of available markers and methodologies. FCSR is focused on addressing the needs of both seasoned flow cytometry users as well as those just discovering the capabilities of flow cytometry tech nology. To this end, FCSR provides monthly training for the instruments for new users and offers support in experimental design and data interpretation following training. Over the current funding period, the FCSR significantly expanded capabilities through the addition of Fortessa and Astrios instruments. In October 2019, the FCSR was awarded an S10 shared instrumentation grant for purchasing a NanoCellect WOLF. Instruments available via the FCSR include a Beckman Coulter MoFlo Astrios (5 lasers and 25 parameters) sorter, a NanoCellect WOLF (1 laser and 5 parameters), a BD Fortessa X-20 (5 lasers and 20 parameters), a BD FACSCanto II analyzer (3 lasers and 10 parameters), a BD FACS Aria cell sorter (3 lasers and 11 parameters), a BD FACSCalibur, and a BD Accuri C6 analyzer (2 lasers and 6 parameters). In addition to the software with each flow cytometer, software is available to analyze multiparameter data to identify and characterize cell subpopulations. DNA histogram analysis software programs are also available for researchers requiring advanced cell cycle analysis. The FCSR is under the directorship of Jason M. Grayson, PhD and has a staff of 1.93 FTEs. To accomplish the goals of the facility, the FCSR has the following Specific Aims: (1) provide cutting-edge analysis instrumentation for flow cytometric analysis; (2) provide multi-parameter cell sorting for user-defined populations; (3) provide instrument training for new and existing users; and (4) provide expertise for experimental design and interpretation of flow cytometry data. From November 1, 2015 to October 16, 2020, the FCSR provided services to 85 users, 60 of whom were WFBCCC members, with 75% supported by peer-reviewed funding. FCSR services were used by investigators representing all four WFBCCC programs and have supported research for 44 grants, including 20 NCI-funded projects, and multiple DoD grants. In the current funding period, 79 publications reported research that utilized the FCSR, including 11 publications with an impact factor greater than 10.