The Duke Cancer Institute Flow Cytometry Shared Resource (DCI-FCSR) was established in 1979 to support investigators conducting basic and clinical cancer research. The DCI-FCSR provides equipment and expertise to allow investigators to analyze and sort complex cell populations. In the Jones building on the Duke main campus, the DCI-FCSR maintains five cell sorters (two Beckman-Coulter Astrios sorters, one BD FACSDiscover S8 Spectral/Imaging sorter, one Sony SH800, one Sony MA900 sorter), and five Flow Cytometry analyzers (two Becton-Dickinson FACSCanto II, one Becton-Dickinson FortessaX20, one Becton-Dickinson FACSymphony A5-SE, one Cytek Aurora). All cell sorters except one Astrios sorter are housed in biosafety cabinets for BSL2/2+ cell sorting. All analyzers and the Sony cell sorters are available for independent use following training by the staff, whereas the Beckman-Coulter and Beckton-Dickinson sorters are staff-operated. The DCI-FCSR also provides a Windows workstation running latest versions of FlowJo and Cytek-SpectroFlo software for post-acquisition data analysis. Experienced DCI-FCSR staff members are available to provide expertise in assay development, assisted data acquisition and cell sorting, data analysis, and to train investigators to independently use facility equipment.