Multiparameter Phenotyping of Human PBMCs Using Mass Cytometry
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.
Key wordsMass cytometry CyTOF Immunophenotyping Panel design
The authors thank Sean Bendall for helpful discussions, and Sheena Gupta and Meena Malipatlolla for contributing example data. Development of this protocol was funded in part by grant 2 U19 AI057229 S4 from the National Institutes of Health.
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