title: Multiparameter Phenotyping of Human PBMCs Using Mass Cytometry.
abstract: The standard for single-cell analysis of phenotype and function in recent decades has been fluorescence flow cytometry. Mass cytometry is a newer technology that uses heavy metal ions, rather than fluorochromes, as labels for probes such as antibodies. The binding of these ion-labeled probes to cells is quantitated by mass spectrometry. This greatly increases the number of phenotypic and functional markers that can be probed simultaneously. Here, we review topics that must be considered when adapting existing flow cytometry panels to mass cytometry analysis. We present a protocol and representative panels for surface phenotyping and intracellular cytokine staining (ICS) assays.
authors.affiliation: Institute for Immunity, Transplantation, and Infection, Stanford University, Stanford, CA, 94305, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Leipold
authors.affiliation: Singapore Immunology Network, Singapore, Singapore.
authors.forename: Evan W
authors.initials: EW
authors.lastname: Newell
authors.affiliation: Institute for Immunity, Transplantation, and Infection, Department of Microbiology and Immunology, Stanford University, Stanford, CA, 94305, USA. maecker@stanford.edu.
authors.forename: Holden T
authors.initials: HT
authors.lastname: Maecker
mesh_terms: D000954:Antigens, Surface / Q000378:metabolism; D015415:Biomarkers; D016207:Cytokines / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D016130:Immunophenotyping / Q000379:methods; D007963:Leukocytes, Mononuclear / Q000378:metabolism*
chemical_list: D000954:Antigens, Surface; D015415:Biomarkers; D016207:Cytokines
keywords: CyTOF; Immunophenotyping; Mass cytometry; Panel design