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

Research output: Contribution to journalJournal article – Annual report year: 2018Researchpeer-review

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DOI

  • Author: Leipold, Michael D.

    Stanford University

  • Author: Obermoser, Gerlinde

    Stanford University

  • Author: Fenwick, Craig

    University of Lausanne

  • Author: Kleinstuber, Katja

    Massachusetts Institute of Technology

  • Author: Rashidi, Narges

    Massachusetts Institute of Technology

  • Author: McNevin, John P.

    Fred Hutchinson Cancer Research Center

  • Author: Nau, Allison N.

    Stanford University

  • Author: Wagar, Lisa E.

    Stanford University

  • Author: Rozot, Virginie

    University of Cape Town

  • Author: Davis, Mark M.

    Stanford University

  • Author: DeRosa, Stephen

    Fred Hutchinson Cancer Research Center

  • Author: Pantaleo, Giuseppe

    University of Lausanne

  • Author: Scriba, Thomas J.

    University of Cape Town

  • Author: Walker, Bruce D.

    Massachusetts Institute of Technology

  • Author: Olsen, Lars R.

    Department of Bio and Health Informatics, Technical University of Denmark, Kemitorvet, 2800, Kgs. Lyngby, Denmark

  • Author: Maecker, Holden T.

    Stanford University

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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. <. 30%. Increased background was seen when using the premixed antibody cocktail aliquots at each site, suggesting that cocktails are best made fresh. Both normalization methods tested performed adequately for normalizing MSI values between centers. Clustering algorithms revealed slight differences between the prestained and the sites-stained samples, due mostly to the increased background of a few antibodies. Therefore, we believe that multicenter mass cytometry assays are feasible.

Original languageEnglish
JournalJournal of Immunological Methods
Volume453
Pages (from-to)37-43
ISSN0022-1759
DOIs
Publication statusPublished - 2018
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • Clustering, CyTOF, Mass cytometry, Multicenter, Normalization, Standardization
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