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Updated: Jun 20, 2026
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An Automated Microscopic Scoring Method for the γ-H2AX Foci Assay in Human Peripheral Blood Lymphocytes
Published on: December 25, 2021
High throughput evaluation of gamma-H2AX
Dane Avondoglio1, Tamalee Scott, Whoon Jong Kil
1Radiation Oncology Branch, National Cancer Institute, National Cancer Institute, Bethesda, Maryland, USA. avondogd@mail.nih.gov
Radiation Oncology (London, England)
|August 26, 2009
Summary
A new assay quantifies DNA double-strand breaks (DSBs) using gamma-H2AX, a marker for radiation damage. This high-throughput method is faster and more reproducible than previous techniques.
Area of Science:
- Molecular Biology
- Radiation Oncology
- Biochemistry
Background:
- DNA double-strand breaks (DSBs) are critical lethal lesions induced by therapeutic radiation.
- Gamma-H2AX foci serve as a recognized biomarker for DSBs.
- Current gamma-H2AX detection assays are labor-intensive and not suitable for high-throughput screening.
Purpose of the Study:
- To develop a novel, high-throughput assay for quantifying gamma-H2AX levels.
- To establish a more efficient and reproducible method for assessing radiation-induced DNA damage.
Main Methods:
- Utilized a high-throughput electrochemiluminescence platform (Mesoscale Discovery Systems).
- Quantified gamma-H2AX levels as a marker for DNA double-strand breaks.
Main Results:
- The developed assay significantly reduces time and labor compared to traditional methods.
- Demonstrated superior intra-assay reproducibility and a wider dynamic range of gamma-H2AX detection.
- Showcased a clear correlation between irradiation dose and gamma-H2AX levels.
Conclusions:
- The new electrochemiluminescence-based assay provides a robust, high-throughput method for quantifying gamma-H2AX.
- This assay has significant potential for preclinical and clinical applications in radiation oncology.
- Offers improved efficiency and reliability for assessing DNA damage response.
