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

Article DOI: 10.3791/62623-v • 08:23 min • December 25th, 2021
December 25th, 2021 •
1, 2, 1, 3, 1, 1, 1, 1, 4, 5, 6, 1
1Radiation Biophysics Division, Nuclear Medicine Department, iThemba LABS, 2Department of Biotechnology, University of the Western Cape, 3Department of Medical Biosciences, University of the Western Cape, 4Division of Medical Physiology, Department of Medicine and Health Sciences, Stellenbosch University, 5Biodosimetry Service, Instituto de Investigaciones Biológicas Clemente Estable and Academic Unit on Radiation Protection, Faculty of Medicine, University of the Republic, 6MetaSystems Hard & Software GmbH

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Summary
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This protocol presents the slide preparation and automated scoring of the γ-H2AX foci assay on peripheral blood lymphocytes. To illustrate the method and sensitivity of the assay, isolated lymphocytes were irradiated in vitro. This automated method of DNA DSB detection is useful for fast and high-throughput biological dosimetry applications.

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Automated Microscopic Scoring γ-H2AX Foci Assay Human Peripheral Blood Lymphocytes Batch Symmetry Applications Radiation Risks DNA Double-strand Breaks Histone Variant H2AX Repair Proteins Microscopy Slide Scanning System Fluorescent Microscope Density Gradient Medium Centrifuge Lymphocytes Counting Hemocytometer
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