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Visualization of Mass Cytometry Signal Background to Enable Optimal Core Panel Customization and Signal Threshold Gating

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Book cover Mass Cytometry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1989))

Abstract

Signal interference or overlap in mass cytometry is minimal compared to flow cytometry but must still be considered for optimal panel design and assay sensitivity. Here we describe a procedure for evaluating signal interference dynamics in the context of a 25-parameter core immunophenotyping panel. Specifically, a mass-minus-many (MMM) approach was used to assess background signals in “empty” or “blank” channels intended for further customization. Through this approach cell type-specific variability in signal background is revealed. Further panel customization can thus be performed with an understanding of cell type and channel-specific background levels to enable rational panel design and the objective delineation of gating thresholds during analysis.

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References

  1. Ornatsky OI, Kinach R, Bandura DR et al (2008) Development of analytical methods for multiplex bio-assay with inductively coupled plasma mass spectrometry. J Anal At Spectrom 23:463–468

    Article  CAS  Google Scholar 

  2. Takahashi C, Au-Yeung A, Fuh F et al (2017) Mass cytometry panel optimization through the designed distribution of signal interference. Cytometry A 91:39–47

    Article  CAS  Google Scholar 

  3. Nicholas KJ, Greenplate AR, Flaherty DK et al (2015) Multiparameter analysis of stimulated human peripheral blood mono-nuclear cells: a comparison of mass and fluorescence cytometry. Cytometry A 89:271–280

    Article  Google Scholar 

  4. Finck R, Simonds EF, Jager A et al (2013) Normalization of mass cytometry data with bead standards. Cytometry A 83(5):483–494. https://doi.org/10.1002/cyto.a.22271

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. OMNI Biomarkers, Development Sciences, Genentech, South San Francisco, CA, USA

    Amelia Au-Yeung, Chikara Takahashi, W. Rodney Mathews & William E. O’Gorman

Authors
  1. Amelia Au-Yeung
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  2. Chikara Takahashi
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  3. W. Rodney Mathews
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  4. William E. O’Gorman
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Corresponding author

Correspondence to William E. O’Gorman .

Editor information

Editors and Affiliations

  1. Discipline of Pathology, The University of Sydney, Camperdown, NSW, Australia

    Helen M. McGuire

  2. Sydney Cytometry Facility, The University of Sydney and Centenary Institute, Camperdown, NSW, Australia

    Thomas M. Ashhurst

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© 2019 Springer Science+Business Media, LLC, part of Springer Nature

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Au-Yeung, A., Takahashi, C., Mathews, W.R., O’Gorman, W.E. (2019). Visualization of Mass Cytometry Signal Background to Enable Optimal Core Panel Customization and Signal Threshold Gating. In: McGuire, H., Ashhurst, T. (eds) Mass Cytometry. Methods in Molecular Biology, vol 1989. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9454-0_3

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  • DOI: https://doi.org/10.1007/978-1-4939-9454-0_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9453-3

  • Online ISBN: 978-1-4939-9454-0

  • eBook Packages: Springer Protocols

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