Investigation of radiation damage in DNA by using atomic force microscopy.
The effect of radiations on supercoiled plasmid DNA has been investigated by using atomic force microscopy (AFM). The DNA molecules were deposited on a substrate and observed by AFM. Alternatively, DNA at different scavenger concentrations was initially exposed to different types of radiations (alpha and X rays) at various doses. After irradiation, fragments (open circular and linearised strands) were observed corresponding to single strand breaks and double strand breaks in DNA. This result indicates the capabilities of AFM for the qualitative detection of strand modifications due to irradiation. The amount of each class of topology enables a quantitative response to be determined for both types of radiation (alpha, X). A value of the radiosensitivity of DNA was obtained as a function of the scavenger concentration. Strong accordance was found between AFM results and those obtained by use of gel electrophoresis. The advantage of AFM in comparison with traditional techniques is the possibility of analysing the radiation effects on one molecule. Indeed, taking the example of alpha particles, it is shown that it is easy to measure the sizes of linear strands by AFM. Such additional or even precise results are difficult to obtain with gel electrophoresis since, in such a case, data are lost through smearing.
Laboratoire de Microanalyses Nucléaires, 16, route de Gray, 25030 Besançon, France. sboichot@netcourrier.com
S
S
Boichot
M
M
Fromm
S
S
Cunniffe
P
P
O'Neill
J C
JC
Labrune
A
A
Chambaudet
E
E
Delain
E
E
Le Cam
D000512:Alpha Particles; D004247:DNA / Q000528:radiation effects* / Q000648:ultrastructure*; D004249:DNA Damage / Q000528:radiation effects*; D004278:DNA, Superhelical / Q000528:radiation effects / Q000648:ultrastructure; D004307:Dose-Response Relationship, Radiation; D018625:Microscopy, Atomic Force; D010957:Plasmids / Q000528:radiation effects* / Q000648:ultrastructure
D004278:DNA, Superhelical; D004247:DNA