Detection in Situ of γ-ray-induced DNA Strand Breaks in Single Cells: Enzymatic Labelling of Free 3′-OH Ends

Author： Fertil B. Modak S. Chavaudra N. Debry H. Meyer F. Malaise E.P.

ISSN： 1362-3095

Source： International Journal of Radiation Biology, Vol.46, Iss.5, 1984-11, pp. : 529-540

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Abstract

We report a procedure allowing the detection and counting of free 3′-OH DNA strand extremities in single cells in situ. Terminal transferase (TdT) catalysed the incorporation of ³H-dGMP into fixed nuclei of human colonic adenocarcinoma cells (HT29), using free 3′-OH ends as initiator. Radioactivity was detected by autoradiography and determined quantitatively with a rapid image-processing system for grain counting. The initiator activity for TdT increases with the dose of γ-rays in the dose range 2·5-20 Gy.

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