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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

Published on: June 26, 2018

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One-Tube Multicolor Flow Cytometry Assay (OTMA) for Comprehensive Immunophenotyping of Peripheral Blood

Anna-Jasmina Donaubauer1, Paul F Rühle1, Ina Becker1

  • 1Department of Radiation Oncology, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|December 13, 2018
PubMed

Insights

This study introduces a one-tube multicolor flow cytometry assay (OTMA) for comprehensive immune status determination. The assay efficiently identifies 20 immune cell subsets and 3 activation markers using minimal blood and preparation time.

Area of Science:

  • Immunology
  • Biotechnology
  • Clinical Diagnostics

Background:

  • Advancements in flow cytometry enable detailed immune profiling.
  • Multicolor immunofluorescence panels are key to assessing immune status.

Purpose of the Study:

  • To present a novel one-tube multicolor flow cytometry assay (OTMA).
  • To enable comprehensive immune status determination.
  • To facilitate translational research in clinical trials.

Main Methods:

  • Developed a 15-antibody, 10-color flow cytometry panel.
  • Utilized 100 μL of EDTA-treated whole blood.
  • Assay preparation time under 40 minutes.

Main Results:

  • Identified 20 distinct, clinically relevant immune cell subsets.
  • Quantified three common immune activation markers.
  • Provided a comprehensive overview of major immune cells.

Conclusions:

  • The OTMA offers an easy and comprehensive method for immune status assessment.
  • The assay is adaptable, requires minimal sample, and is suitable for translational research.
  • This method supports clinical trials by providing detailed immune cell counts and activation states.

Keywords:
Absolute cell countAntibodyGating strategyHuman whole-bloodImmune statusImmunophenotypingMulticolor flow cytometryOne-tube measurement

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