Biophysical modeling of fragment length distributions of DNA plasmids after X and heavy-ion irradiation analyzed by atomic force microscopy.

The investigation of fragment length distributions of plasmid DNA gives insight into the influence of localized energy distribution on the induction of DNA damage, particularly the clustering of double-strand breaks. We present an approach that determines the fragment length distributions of plasmid DNA after heavy-ion irradiation by using the Local Effect Model. We find a good agreement of our simulations with experimental fragment distributions derived from atomic force microscopy (AFM) studies by including experimental constraints typical for AFM. Our calculations reveal that by comparing the fragmentation in terms of fluence, we can uniquely distinguish the effect of different radiation qualities. For very high-LET irradiation using nickel or uranium ions, no difference between their fragment distributions can be expected for the same dose level. However, for carbon ions with an intermediate LET, the fragmentation pattern differs from the distribution for very high-LET particles. The results of the model calculations can be used to determine the optimal experimental parameters for a demonstration of the influence of track structure on primary radiation damage. Additionally, we compare the results of our model for two different plasmid geometries.

Direct visualisation of heavy ion induced DNA fragmentation using atomic force microscopy.

Atomic Force Microscopy of phiX174 plasmids irradiated in vitro was used to visualise the DNA fragmentation induced by heavy ions and to compare it to the fragmentation pattern obtained after X-irradiation. Fragment distributions induced by low ion fuences were found to be much more shifted towards small fragment sizes than the distributions obtained after corresponding doses of X-rays. The average fragment length was found to be significantly smaller than the full plasmid length even for single ion traversals.

SFM studies of carbon ion induced damages in plasmid DNA.

In this study we present for the first time detailed scanning force microscopy (SFM) investigations of carbon ion induced damages in plasmid DNA in order to obtain information about the biological effectiveness of particle radiation. For this purpose, we have combined SFM and gel electrophoresis measurements in a dose range between D = 0 Gy and 5000 Gy. After irradiation with C ions, the percentage of double-strand breaks (DSBs) increases drastically, i.e. from initially 0% for D = 0 Gy to 38% for D = 5000 Gy. Increasing the dose over the total range is accompanied by a shortening of the average fragment length from L = 1100 nm to L = 575 nm. In addition to our experiments, the average numbers of induced DSBs per irradiated plasmid and per broken plasmid have been calculated from the SFM measurements. The most important among the numerous results is that a significant amount of plasmids has suffered more than two DSBs for all applied doses, indicating multiple DSBs. The number of DSBs per broken plasmid increases from approximately 1.7 after irradiation with a dose of D = 250 Gy to 3.2 after exposure to the highest dose of D = 5000 Gy. The results provide experimental data for the spatially correlated production of DSBs after carbon irradiation, that are relevant to the understanding of its biological effectiveness.