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Updated: Aug 2, 2025

Cell Cycle-specific Measurement of γH2AX and Apoptosis After Genotoxic Stress by Flow Cytometry

Cell Cycle-specific Measurement of γH2AX and Apoptosis After Genotoxic Stress by Flow Cytometry

Published on: September 1, 2019

High-Throughput γ-H2AX Assay Using Imaging Flow Cytometry.

Younghyun Lee^1,2, Qi Wang^3,4, Ki Moon Seong¹

¹Laboratory of Biological Dosimetry, National Radiation Emergency Medical Center, Korea Institute of Radiological and Medical Sciences, Seoul, Republic of Korea.

Methods in Molecular Biology (Clifton, N.J.)

|April 19, 2023

Summary

This summary is machine-generated.

A new high-throughput gamma-H2AX assay using imaging flow cytometry allows rapid, reliable detection of radiation-induced DNA damage. This method is suitable for large-scale radiation biodosimetry and epidemiological studies.

Keywords:

Double-strand break High-throughput platform Imaging flow cytometry Radiation γ-H2AX

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Last Updated: Aug 2, 2025

Cell Cycle-specific Measurement of γH2AX and Apoptosis After Genotoxic Stress by Flow Cytometry

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Published on: September 1, 2019

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An Automated Microscopic Scoring Method for the γ-H2AX Foci Assay in Human Peripheral Blood Lymphocytes

An Automated Microscopic Scoring Method for the γ-H2AX Foci Assay in Human Peripheral Blood Lymphocytes

Published on: December 25, 2021

Area of Science:

Radiation biology
Molecular biology
Biotechnology

Background:

The gamma-H2AX assay is a standard method for detecting DNA double-strand breaks caused by radiation.
Manual analysis of gamma-H2AX foci is time-consuming and not feasible for high-throughput screening.

Purpose of the Study:

To develop a high-throughput gamma-H2AX assay for efficient radiation biodosimetry.
To adapt the assay for large-scale screening in mass casualty events and epidemiological studies.

Main Methods:

Developed a high-throughput gamma-H2AX assay utilizing imaging flow cytometry.
Employed automated image acquisition with ImageStream®X and data analysis using IDEAS® software.
Utilized small blood volumes processed in a 96-tube format for sample preparation.

Main Results:

The assay enables rapid analysis of thousands of cells from small blood volumes.
Accurate and reliable quantitative measurements of gamma-H2AX foci and mean fluorescence levels were achieved.
Demonstrated suitability for high-throughput screening of radiation-induced DNA damage.

Conclusions:

The developed high-throughput gamma-H2AX assay is a valuable tool for radiation biodosimetry in mass casualty incidents.
This method can support large-scale molecular epidemiological studies and personalized radiotherapy.
Offers a significant advancement over conventional manual gamma-H2AX assays for efficiency and scale.