Multi-site reproducibility of a human immunophenotyping assay in whole blood and peripheral blood mononuclear cells preparations using CyTOF technology coupled with Maxpar Pathsetter, an automated data analysis system

Cytometry B Clin Cytom. 2020 Mar;98(2):146-160. doi: 10.1002/cyto.b.21858. Epub 2019 Nov 23.


High-dimensional mass cytometry data potentially enable a comprehensive characterization of immune cells. In order to positively affect clinical trials and translational clinical research, this advanced technology needs to demonstrate a high reproducibility of results across multiple sites for both peripheral blood mononuclear cells (PBMC) and whole blood preparations. A dry 30-marker broad immunophenotyping panel and customized automated analysis software were recently engineered and are commercially available as the Fluidigm® Maxpar® Direct™ Immune Profiling Assay™. In this study, seven sites received whole blood and six sites received PBMC samples from single donors over a 2-week interval. Each site labeled replicate samples and acquired data on Helios™ instruments using an assay-specific acquisition template. All acquired sample files were then automatically analyzed by Maxpar Pathsetter™ software. A cleanup step eliminated debris, dead cells, aggregates, and normalization beads. The second step automatically enumerated 37 immune cell populations and performed label intensity assessments on all 30 markers. The inter-site reproducibility of the 37 quantified cell populations had consistent population frequencies, with an average %CV of 14.4% for whole blood and 17.7% for PBMC. The dry reagent coupled with automated data analysis is not only convenient but also provides a high degree of reproducibility within and among multiple test sites resulting in a comprehensive yet practical solution for deep immune phenotyping.

Keywords: cytometry automation; cytometry standardization; kits; percentage precision.

MeSH terms

  • Automation, Laboratory / instrumentation
  • Automation, Laboratory / methods
  • Automation, Laboratory / standards
  • Blood Cells / cytology*
  • Canada
  • Data Analysis
  • Flow Cytometry* / instrumentation
  • Flow Cytometry* / methods
  • Flow Cytometry* / standards
  • Humans
  • Immunophenotyping* / instrumentation
  • Immunophenotyping* / methods
  • Immunophenotyping* / standards
  • Laboratory Proficiency Testing
  • Leukocytes, Mononuclear / cytology
  • Pattern Recognition, Automated / methods
  • Pattern Recognition, Automated / standards
  • Reference Standards
  • Reproducibility of Results
  • United States