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Related Experiment Video

Updated: Mar 12, 2026

Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
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Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry

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Optimization of mass cytometry sample cryopreservation after staining.

Hermi R Sumatoh1, Karen Wei Weng Teng1, Yang Cheng1,2

  • 1Agency for Science, Technology and Research (A*STAR), Singapore Immunology Network (SIgN), Singapore, 138648, Singapore.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|November 1, 2016
PubMed
Summary

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Cryopreserving stained mass cytometry samples offers a practical solution to technical challenges. Freezing samples in 10% DMSO in FBS effectively preserves cellular markers for up to 4 weeks.

Area of Science:

  • Single-cell analysis
  • Mass cytometry
  • Immunophenotyping

Background:

  • Mass cytometry enables deep assessment of cellular diversity.
  • Technical hurdles in mass cytometry include equipment breakdown and large-scale batch sampling.
  • Current sample handling approaches present limitations, especially for precious samples.

Purpose of the Study:

  • To evaluate protocols for cryopreserving stained and fixed mass cytometry samples.
  • To compare cryopreservation with standard refrigeration for up to 4 weeks.
  • To assess the impact of freezing on cellular viability and marker signal intensity.

Main Methods:

  • Two cryopreservation protocols were evaluated for stained and fixed cellular samples.
  • Standard sample refrigeration in staining buffer served as a control.
Keywords:
cellular surface markersfreezingintracellular stainingmass cytometry

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  • A comprehensive human T cell panel was used to assess phenotypic and functional markers over 4 weeks.

    • Main Results:

    • Cellular viability, DNA intercalator, and barcode staining were minimally affected by freezing compared to refrigeration.
    • Cell surface marker and receptor signal intensities remained largely uncompromised after freezing.
    • Intracellular cytokine staining showed decreased signal intensity post-freezing, particularly with a methanol-based protocol.

    Conclusions:

    • Cryopreservation of stained mass cytometry samples is a practical and efficient preservation method.
    • Freezing samples in 10% DMSO in FBS is recommended for preserving stained samples.
    • This approach alleviates technical limitations associated with current sample-handling methods.