Comparison of CyTOF assays across sites: Results of a six-center pilot study

For more than five years, high-dimensional mass cytometry has been employed to study immunology. However, these studies have typically been performed in one laboratory on one or few instruments. We present the results of a six-center study using healthy control human peripheral blood mononuclear cells (PBMCs) and commercially available reagents to test the intra-site and inter-site variation of mass cytometers and operators. We used prestained controls generated by the primary center as a reference to compare against samples stained at each individual center. Data were analyzed at the primary center, including investigating the effects of two normalization methods. All six sites performed similarly, with CVs for both Frequency of Parent and median signal intensity (MSI) values
Highlights • A multicenter mass cytometry study was performed among six international centers. • A premixed antibody cocktail resulted in higher background for a few antibodies. • Manual gating and clustering algorithms demonstrate that all sites were adequately consistent in performance.