Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells

Abstract

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.