title: Multiparameter analysis of stimulated human peripheral blood mononuclear cells: A comparison of mass and fluorescence cytometry.
abstract: Mass and fluorescence cytometry are quantitative single cell flow cytometry approaches that are powerful tools for characterizing diverse tissues and cellular systems. Here mass cytometry was directly compared with fluorescence cytometry by studying phenotypes of healthy human peripheral blood mononuclear cells (PBMC) in the context of superantigen stimulation. One mass cytometry panel and five fluorescence cytometry panels were used to measure 20 well-established lymphocyte markers of memory and activation. Comparable frequencies of both common and rare cell subpopulations were observed with fluorescence and mass cytometry using biaxial gating. The unsupervised high-dimensional analysis tool viSNE was then used to analyze data sets generated from both mass and fluorescence cytometry. viSNE analysis effectively characterized PBMC using eight features per cell and identified similar frequencies of activated CD4+ T cells with both technologies. These results suggest combinations of unsupervised analysis programs and extended multiparameter cytometry will be indispensable tools for detecting perturbations in protein expression in both health and disease.
authors.affiliation: Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee.
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Nicholas
authors.affiliation: Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee.|Department of Cancer Biology and Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, Tennessee.
authors.forename: Allison R
authors.initials: AR
authors.lastname: Greenplate
authors.affiliation: Flow Cytometry Shared Resource, Vanderbilt University School of Medicine, Nashville, Tennessee.
authors.forename: David K
authors.initials: DK
authors.lastname: Flaherty
authors.affiliation: Flow Cytometry Shared Resource, Vanderbilt University School of Medicine, Nashville, Tennessee.
authors.forename: Brittany K
authors.initials: BK
authors.lastname: Matlock
authors.affiliation: Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, Tennessee.
authors.forename: Juan San
authors.initials: JS
authors.lastname: Juan
authors.affiliation: Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, Tennessee.
authors.forename: Rita M
authors.initials: RM
authors.lastname: Smith
authors.affiliation: Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee.|Department of Cancer Biology and Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, Tennessee.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Irish
authors.affiliation: Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee.|Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, Tennessee.
authors.forename: Spyros A
authors.initials: SA
authors.lastname: Kalams
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D005434:Flow Cytometry / Q000592:standards*; D015870:Gene Expression; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D028581:Lanthanoid Series Elements / Q000032:analysis; D007963:Leukocytes, Mononuclear / Q000145:classification / Q000166:cytology* / Q000276:immunology; D008213:Lymphocyte Activation; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods / Q000592:standards*; D015999:Multivariate Analysis
chemical_list: D015703:Antigens, CD; D028581:Lanthanoid Series Elements
keywords: CyTOF; T cells; fluorescence cytometry; human PBMC; mass cytometry; viSNE