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Related Experiment Video

Updated: Jul 20, 2025

An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
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Application of the Cytokinesis-Block Micronucleus Assay for High-Dose Exposures Using Imaging Flow Cytometry.

Lindsay A Beaton-Green1, Jessica M Mayenburg1, Leonora Marro1

  • 1Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.

Cytogenetic and Genome Research
|August 1, 2023
PubMed
Summary

The cytokinesis-block micronucleus assay, adapted for imaging flow cytometry, automates radiation damage assessment in human cells. This enhanced method accurately estimates radiation doses up to 10 Gy, improving triage dosimetry efficiency.

Keywords:
BiodosimetryCytokinesis‐block micronucleus assayEmergency responseImaging flow cytometryMulti‐parametric analysisRadiation

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Area of Science:

  • Biomedical sciences
  • Radiation biology
  • Cellular toxicology

Background:

  • The cytokinesis-block micronucleus assay is a standard method for evaluating radiation-induced genetic damage.
  • Adapting this assay to imaging flow cytometry (IFC) enables automated, high-throughput cell analysis without manual slide preparation.

Purpose of the Study:

  • To develop and validate a multi-parametric analysis for the IFC-based micronucleus assay.
  • To extend the dose estimation range of the assay up to 10 Gy for improved triage dosimetry.
  • To confirm the functional equivalence of manual and automated sample acquisition methods.

Main Methods:

  • Human blood samples were irradiated, cultured, stained, and analyzed using IFC with both manual and 96-well plate automated acquisition.
  • Multi-parametric image features were extracted for each cell.
  • Calibration curves were generated using quadratic random coefficient and logistic discriminant models.
  • Dose estimates were validated using blinded samples and assessed for accuracy via relative bias and mean square error.

Main Results:

  • The developed multi-parametric analysis successfully generated dose calibration curves up to 10 Gy.
  • Automated and manual acquisition methods yielded functionally identical results.
  • Dose estimation accuracy was sufficient for triage dosimetry (>90% within 1 Gy for lower doses, ~50% for higher doses).
  • The lowest accuracy occurred between 5-6 Gy due to minimal change in micronuclei frequency with dose.

Conclusions:

  • The novel multi-parametric analysis significantly enhances the utility of the IFC micronucleus assay for radiation dose assessment.
  • The assay's extended range and validated automated acquisition improve efficiency and accuracy in triage dosimetry.
  • This method provides a robust tool for quantifying radiation-induced genetic damage in human cells.