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Acquisition, Processing, and Quality Control of Mass Cytometry Data

Brian H Lee1, Adeeb H Rahman2,3

  • 1Human Immune Monitoring Center, Icahn School of Medicine at Mt. Sinai, New York, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|May 12, 2019
PubMed

Insights

Mass cytometry, a powerful technique for immune cell analysis, enhances single-cell characterization. This guide details optimal data acquisition and processing for mass cytometry to improve cell population analysis.

Area of Science:

  • Immunology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Mass cytometry offers high-dimensional immune cell profiling by combining mass spectrometry and flow cytometry principles.
  • It utilizes isotopically conjugated antibodies to overcome spectral overlap limitations inherent in traditional flow cytometry.
  • This enables deeper single-cell characterization of complex biological samples with an expanded panel of cellular markers.

Purpose of the Study:

  • To provide an overview of optimal data acquisition strategies for mass cytometry.
  • To outline essential data processing techniques for mass cytometry experiments.
  • To facilitate the accurate analysis and characterization of immune cell populations.

Main Methods:

  • Detailed explanation of mass cytometry principles and instrumentation.
  • Guidance on selecting and conjugating metal isotopes to antibodies.
  • Best practices for sample preparation and instrument calibration for mass cytometry.

Main Results:

  • Established protocols for high-quality mass cytometry data acquisition.
  • Demonstrated methods for effective data preprocessing, including debarcoding and normalization.
  • Improved accuracy in identifying and quantifying immune cell subsets.

Conclusions:

  • Mass cytometry provides unparalleled depth in single-cell immune profiling.
  • Adherence to optimal data acquisition and processing is crucial for reliable results.
  • This approach significantly advances the characterization of complex immune responses.

Keywords:
BarcodingData acquisitionData processingMass cytometryNormalization

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