Measurement of DNA damage in mammalian cells using flow cytometry

Journal Article · Wed Apr 01 00:00:00 EST 1987 · Radiat. Res.; (United States)

DOI:https://doi.org/10.2307/3576888· OSTI ID:6788087

Milner, A E; Vaughan, A T; Clark, I P

A technique for the detection of DNA damage induced by radiation insult has been developed. Cells were lysed with a buffer containing 2 M sodium chloride to release the DNA in a supercoiled form, the nucleoid. These were stained with the DNA intercalating dye, ethidium bromide, and exposed to laser light within a flow cytometer. Scattered and fluorescent light was analyzed from the laser/nucleoid interaction following irradiation of viable cells with gamma rays. The addition of ethidium bromide to prepared nucleoids caused a reduction in scattered light due to condensation of the nucleoid. Irradiation of cells prior to nucleoid production and ethidium bromide treatment restricted this condensation and produced a dose-dependent increase in laser scatter. Nucleoids derived from human lymphocytes showed enhanced light scatter from 5 Gy, compared to Chinese hamster ovary (CHO) fibroblasts where doses above 10 Gy were required. Up to 30 Gy CHO nucleoids showed a dose-dependent reduction in the ethidium bromide fluorescence. This technique allows detection of altered light scattering and fluorescent behavior of nucleoids after cellular irradiation; these may be related to structural changes within the nucleus induced by the radiation. The use of flow cytometry compared to other methods allows a rapid analysis of nuclear damage within individual cells.

• Research Organization: Univ. of Birmingham, Edgbaston, England
• OSTI ID: 6788087
• Journal Information: Radiat. Res.; (United States), Vol. 1
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
FIBROBLASTS
GENETIC RADIATION EFFECTS
LASER RADIATION
GENETIC EFFECTS
LYMPHOCYTES
CELL FLOW SYSTEMS
CHO CELLS
DNA
DOSE-RESPONSE RELATIONSHIPS
GAMMA RADIATION
HAMSTERS
IN VITRO
STAINS
ANIMAL CELLS
ANIMALS
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
BIOLOGICAL RADIATION EFFECTS
BLOOD
BLOOD CELLS
BODY FLUIDS
CONNECTIVE TISSUE CELLS
ELECTROMAGNETIC RADIATION
IONIZING RADIATIONS
LEUKOCYTES
MAMMALS
MATERIALS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
RADIATION EFFECTS
RADIATIONS
RODENTS
SOMATIC CELLS
VERTEBRATES
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture
560400 - Other Environmental Pollutant Effects