All antibodies were obtained from Fluidigm Maxpar Human Peripheral Blood Basic I Phenotyping Panel Kit (Catalog #: 201302). For cell staining protocols, Cell Staining Buffer (CSB) (Catalog #: 201068), Intercalator-Ir (Catalog #: 201192A, 125 μM), water (Catalog #: 201069), and Cell Acquisition Solution (CAS) (Catalog # 201240) were all acquired from Fluidigm (San Francisco, Ca). Pierce™ 16% formaldehyde (Catalog #: 28908) was purchased from Thermo Fisher Scientific (Waltham MA) to make a working concentration of 1.6% in PBS. Reagents for sample analysis on the Helios mass cytometer included Tuning Solution (Catalog #: 201072), EQ™ Four Element Calibration Beads (EQ4, Catalog #: 201078), and Washing Solution (Catalog #: 201070) were also acquired from Fluidigm. Five Antibody Binding Capacity (ABC) calibration beads were synthesized using dispersion polymerization, independently characterized for the number of metal atoms per bead [44 (link)] and combined for use in mass cytometry. The five beads feature a roughly equivalent amount of Ce, with zero and then logarithmically increasing amounts of La, Eu, Ho, and Lu.
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