Spectral Flow Cytometry Analysis of Seven Healthy Adults

Overview This data is part of a larger project that is ongoing. This subset includes eight manually gated cell populations from seven healthy, non-pregnant adult patients. Analysis was performed on a Cytek Aurora. Methods Peripheral blood was obtained from healthy, non-pregnant adult donors. All human samples used for this research were de-identified, with discarded clinical material and no patient or clinical information available for analysis. Whole blood from each donor was diluted 1:1 in 1x PBS to a final volume of 50 mL and 25 mL aliquots were layered on 12 mL Ficoll gradient. After gradient centrifugation (20 min, 2000 rpm), lymphocytes were isolated from the Ficoll interface and washed once in RPMI containing pen/strep and 10% FBS and directly stained for analysis on a 5-laser Cytek Aurora spectral flow cytometer. Antibodies and isotype controls used for flow cytometric analysis are listed in the Panel.pdf document. For surface staining, cells were stained for 30 min in RPMI containing pen/strep and 10% FBS. For intracellular staining, cells were fixed and permeabilized using the eBiosciences FOXP3 kit (eBiosciences). Analysis was performed on a Cytek Aurora. All flow cytometric data were acquired using equipment maintained by the Research Flow Cytometry Core in the Division of Rheumatology at Cincinnati Children’s Hospital Medical Center. Additional Information Tursi AR, Lages CS, Quayle K, Koenig ZT, Loni R, Eswar S, Cobeña-Reyes J, Thornton S, Tilburgs T, Andorf S. Automated descriptive cell type naming in flow and mass cytometry with CytoPheno. Sci Rep. 2025 Jul 23;15(1):26750. doi: 10.1038/s41598-025-12153-w. PMID: 40702123; PMCID: PMC12287297.