Articles by William E. O'Gorman in JoVE
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry Ryan M. Baxter*1, Daniel S. Kong*1, Josselyn E. Garcia-Perez1, William E. O'Gorman2, Elena W.Y. Hsieh1,3 1Department of Immunology and Microbiology, University of Colorado School of Medicine, 2OMNI Biomarkers, Development Sciences, Genentech, 3Department of Pediatrics, Division of Allergy and Immunology, University of Colorado School of Medicine Here we describe a single-cell proteomic approach to evaluate immune phenotypic and functional (intracellular cytokine induction) alterations in peripheral whole blood samples, analyzed via mass cytometry.
Other articles by William E. O'Gorman on PubMed
Mass Cytometry Panel Optimization Through the Designed Distribution of Signal Interference Cytometry. Part A : the Journal of the International Society for Analytical Cytology. | Pubmed ID: 27632576 Mass cytometry is capable of measuring more than 40 distinct proteins on individual cells making it a promising technology for innovating biomarker discovery. However, in order for this potential to be fully realized, best practices in panel design need to be further defined in order to achieve consistency and reproducibility in data analysis. Of particular importance are controls that reveal, and panel design principles that mitigate the effects of signal interference or overlap. We observed a disparity between the staining profiles of two noncompeting anti- integrin β7 mAbs and hypothesized that signal interference was responsible. A mass-minus-one (MMO) control was applied and demonstrated that signal overlap caused the perceived interclonal discrepancy in β7 expression. Panel redesign in consideration of mass-cytometry specific interference dynamics dramatically improved concordance between both mAbs by redistributing background signals caused by overlap. These studies visualize how signal overlap can complicate mass cytometry data interpretation and demonstrate how the rational distribution of interference can greatly improve panel design and data quality. © 2016 International Society for Advancement of Cytometry.