PROJECT SUMMARY / ABSTRACT Since its inception in 1988, the Flow Cytometry Laboratory (FLOW) has served as the only comprehensive shared resource on the UW-Madison campus for multiparameter flow cytometry, cell sorting and imaging cell cytometry. The mission of FLOW is to operate a cutting-edge facility that provides UWCCC researchers the infrastructure, technical expertise, and educational support required for fluorescence-based single cell analysis and isolation that advances basic, translational, and clinical research. FLOW manages one 67-parameter Cytek Aurora, five 16-parameter ThermoFisher Attune, one 20-parameter BD LSRII, and one 20-parameter BD LSR Fortessa analytical cytometers; three BD FACSAria and one Sony MA900 cell sorters; and a Luminex ImageStreamX MarkII imaging cytometer. Strong institutional support and funds from NIH S10 Shared Instrumentation Grants have allowed FLOW to purchase and maintain state-of-the-art instruments, computers, and software needed for data acquisition and analysis. FLOW operates facilities at two locations: 1) our main laboratory in the Wisconsin Institute for Medical Research (WIMR), which is located on West Campus and houses the majority of UWCCC researchers; and 2) a satellite facility in the UW Biotechnology Center (UWBC), which is located on Central Campus near our remaining members and contiguous to facilities that provide DNA, RNA, and single-cell next-generation sequencing to characterize freshly sorted single cells and cell populations. FLOW partners with the Department of Pediatrics and the UWCCC Small Animal Imaging and Radiotherapy Facility (SAIRF) to operate cytometers dedicated primarily to studies of pediatric cancers and analysis of radioactive samples, respectively. The latter partnership was established to support targeted radionuclide therapy of pediatric tumors in combination with immunotherapy. Dr. Manish Patankar (TM), a UWCCC member and ovarian cancer researcher, serves as faculty leader. Ms. Dagna Sheerar has managed day-to-day operations since 2011. Ms. Sheerar and her 6 staff members possess over 80 years of collective experience in cytometry. The specific aims for FLOW are to: 1) evaluate new technologies and develop business plans to procure and maintain new instruments allowing users access to cutting-edge approaches to single cell analytics and sorting; 2) provide expert consultation and education services for single cell analytics and cell sorting ; and 3) organize user groups and develop data analysis support in collaboration with the Cancer Informatics Shared Resource for the increasingly complex data sets generated by multi-parameter cytometers and sorters. During the current CCSG funding cycle, FLOW served 109 unique UWCCC program members across all six programs who collectively hold/held 445 cancer relevant research grants, including 118 awards from the NCI, and contributed to 169 cancer relevant UWCCC member publications. Survey data indicate a high level of user sati...