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2 INSERM U1223, Paris, France
3 Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France
4 Flow Cytometry Platfrom, Cytometry and Biomarkers UtechS, Center for Translational Science, Pasteur Institute, Paris, France
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2 INSERM U1223, Paris, France
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Related Experiment Video

Updated: Jan 19, 2026

Automation of the Micronucleus Assay Using Imaging Flow Cytometry and Artificial Intelligence
09:11

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Guidelines for Gating Flow Cytometry Data for Immunological Assays.

Janet Staats1, Anagha Divekar2, J Philip McCoy3

  • 1Duke Immune Profiling Core, Duke University Medical Center, Durham, NC, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 16, 2019
PubMed
Summary
This summary is machine-generated.

Flow cytometry gating, the manual selection of cell populations, lacks standardized guidelines. This study provides comprehensive recommendations for consistent and accurate cell subpopulation analysis in immunological assays.

Keywords:
AnalysisFlow cytometryGating

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Flow cytometry gating is crucial for analyzing cell subpopulations.
  • Manual gating relies on expert knowledge but suffers from inter-observer variability.
  • Existing guidelines are assay-specific, lacking comprehensive coverage for diverse immunological assays.

Purpose of the Study:

  • To establish comprehensive guidelines for flow cytometry gating across various immunological assays.
  • To address the lack of standardization in manual gating procedures.
  • To improve the consistency and accuracy of cell subpopulation analysis.

Main Methods:

  • Development of general gating guidelines based on common cell characteristics.
  • Inclusion of example data from academic, government, and industry sources.
  • Focus on fundamental gating strategies applicable to diverse immunological assays.

Main Results:

  • Presentation of recommendations for the most common and critical gating strategies.
  • Demonstration of gating principles using representative flow cytometry data.
  • Consensus-based approach to guide gating practices.

Conclusions:

  • Standardized flow cytometry gating guidelines are essential for reliable immunological assay results.
  • These guidelines aim to reduce variability and enhance reproducibility in cell analysis.
  • The recommendations are valuable for both novice and experienced researchers in flow cytometry.