Ostling O, Johanson K J
Biochem Biophys Res Commun. 1984 Aug 30;123(1):291-8. doi: 10.1016/0006-291x(84)90411-x.
Mammalian cells were after irradiation suspended in melted agarose, and casted on microscope slides. The slides were after gelling at 0 degree C immersed in a neutral detergent solution which lysed the cells. A weak electric field (5 V/cm) was then applied over the gel for 5 minutes. The DNA in the gel was stained with the fluorescent dye acridine orange and gives a green emission in a microscope photometer. DNA had migrated towards the anode and this migration was more pronounced in irradiated than in control cells. The differences in migration pattern were quantitatively measured. The lower detection limit was below 0.5 Gy and a plateau in the dose-effect curve was reached at about 3 Gy. In repair experiments residual DNA damage could be observed after postirradiation incubation for 60 minutes. The advantages of the method is: no radioactive labelling and only a few number of cells is required.
将哺乳动物细胞在辐照后悬浮于融化的琼脂糖中，并浇铸在显微镜载玻片上。载玻片在0℃凝胶化后，浸入使细胞裂解的中性去污剂溶液中。然后在凝胶上施加弱电场（5V/cm）5分钟。凝胶中的DNA用荧光染料吖啶橙染色，并在显微镜光度计中发出绿色荧光。DNA向阳极迁移，且这种迁移在受辐照细胞中比在对照细胞中更明显。对迁移模式的差异进行了定量测量。检测下限低于0.5Gy，剂量效应曲线在约3Gy处达到平台期。在修复实验中，辐照后孵育60分钟后可观察到残留的DNA损伤。该方法的优点是：无需放射性标记，且仅需少量细胞。
