High-Parameter Immune Profiling with CyTOF

  • Bita Sahaf
  • Adeeb Rahman
  • Holden T. MaeckerEmail author
  • Sean C. Bendall
Part of the Methods in Molecular Biology book series (MIMB, volume 2055)


Mass cytometry, or CyTOF, is a useful technology for high-parameter single-cell phenotyping, especially from suspension cells such as blood or PBMC. It is particularly appealing to monitor the systemic immune changes that could accompany cancer immunotherapy. Here we present a reference panel for identification of all major immune cell populations, with flexibility for addition of trial-specific markers. We also describe best-practice measures for minimizing and tracking batch variability. These include: sample barcoding, use of spiked-in reference cells, and lyophilization of the antibody cocktail. Our protocol assumes the use of cryopreserved PBMC, both for convenience of batching samples and for maximum comparability across patients and time points. Finally, we show an option for automated analysis using the Astrolabe platform (Astrolabe Diagnostics, Inc.).

Key words

Mass cytometry Differential analysis Phenotypic markers Subpopulation Multiplexed 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Bita Sahaf
    • 1
  • Adeeb Rahman
    • 2
  • Holden T. Maecker
    • 3
    Email author
  • Sean C. Bendall
    • 4
  1. 1.PICI Cancer Correlative Science Unit, Cancer InstituteStanford University School of MedicineStanfordUSA
  2. 2.Department of Genetics and Genomic SciencesIcahn School of Medicine at Mount SinaiNew YorkUSA
  3. 3.Institute for Immunity, Transplantation, and InfectionStanford University School of MedicineStanfordUSA
  4. 4.Department of PathologyStanford University School of MedicineStanfordUSA

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