FLOW CYTOMETRY & CELL SORTING (FCCS) SHARED RESOURCE: SUMMARY The mission of the Flow Cytometry & Cell Sorting (FCCS) Shared Resource is to provide the Hollings Cancer Center (HCC) research community with an integrated platform to sort and analyze cells and evaluate pre- clinical and clinical experimental data in a cell-specific manner. Under the leadership of Amanda C. LaRue, PhD (CBI), FCCS offers comprehensive high-speed cell sorting services, high-dimensional single-cell analyses, the development of new methodologies, and an educational platform to further basic and translational research. Flow cytometric techniques allow for rapid phenotypic and functional analysis of biological processes and isolation of single cells, methodologies that are critical for detailed analyses of cell types contributing to the complex processes of cancer initiation, metastasis, and progression. Immune monitoring services evaluate patients' immune responses and provide valuable data needed for preclinical and basic laboratory investigations in animals or tissue culture. These methodologies have provided essential insights into how cancer evolves, opening new opportunities for therapeutic interventions. This is accomplished by providing assisted and unassisted flow cytometry analysis, consultation on experimental design, sample preparation, data analysis, and preparation, along with hands-on training in flow cytometric technology and analytical equipment. Due to substantive new equipment acquisition, multi-parametric, high-dimensional analyses have expanded during this cycle. FCCS added a Hyperion to the CyTOF System for imaging mass cytometry that allows antibody-based detection and analysis of up to 135 simultaneous targets. FCCS also added a Beckman Coulter CytoFLEX LX and a Cytek Northern Lights, both high-dimensional “next-generation” analyzers. The primary sorter, a BD FACSAria II, has also been upgraded to a BD FACSAria III. To meet the educational mission of FCCS, a formal, comprehensive training program, Ready, Set, Flow, was developed during the last review period. During the current cycle, FCCS was utilized by 70 HCC members and provided key data and insights to support the success of 112 peer-reviewed publications, including work published in high-impact journals such as Nature Med, JCI, Cell Reports, Cell Mol Immunol, Cancer Res, Cancer Immunol, and JBC. In summary, FCCS advances, supports, and provides critical, cutting-edge research tools and methodologies to the HCC community.