MICROELECTROPHORETIC STUDY OF RADIATION-INDUCED DNA DAMAGES IN INDIVIDUAL MAMMALIAN-CELLS
MICROELECTROPHORETIC STUDY OF RADIATION-INDUCED DNA DAMAGES IN INDIVIDUAL MAMMALIAN-CELLS
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DOI：
10.1016/0006-291x(84)90411-x
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发表时间：
1984-01-01
影响因子：
3.1
通讯作者：
JOHANSON, KJ
中科院分区：
文献类型：
--
作者：
OSTLING, O;JOHANSON, KJ
照射后，将哺乳动物细胞悬浮在熔化的琼脂糖中，并浇铸在显微镜载玻片上。载玻片在0 ℃下胶凝后，C浸入中性洗涤剂溶液中，溶解细胞。然后在凝胶上施加弱电场（5 V/cm）5 min。凝胶中的DNA用荧光染料吖啶橙子染色，在显微镜光度计中发出绿色光。DNA已经向阳极迁移，并且这种迁移在辐照细胞中比在对照细胞中更明显。定量测量了迁移模式的差异。检测下限低于0.5戈伊[gray]，剂量效应曲线在约3戈伊时达到平台。在修复实验中，辐射后孵育60分钟后可以观察到残留的DNA损伤。该方法的优点是不需要放射性标记，只需要少量的细胞。
Mammalian cells were suspended in melted agarose after irradiation 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 min. 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 [gray] 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 min. The advantages of the method is no radioactive labeling and only a few number of cells is required.