Nanoscale detection of ionizing radiation damage to DNA by atomic force microscopy.
The detection and quantification of ionizing radiation damage to DNA at a single-molecule level by atomic force microscopy (AFM) is reported. The DNA damage-detection technique combining supercoiled plasmid relaxation assay with AFM imaging is a direct and quantitative approach to detect gamma-ray-induced single- and double-strand breaks in DNA, and its accuracy and reliability are validated through a comparison with traditional agarose gel electrophoresis. In addition, the dependence of radiation-induced single-strand breaks on plasmid size and concentration at a single-molecule level in a low-dose (1 Gy) and low-concentration range (0.01 ng microL(-1)-10 ng microL(-1)) is investigated using the AFM-based damage-detection assay. The results clearly show that the number of single-strand breaks per DNA molecule is linearly proportional to the plasmid size and inversely correlated to the DNA concentration. This assay can also efficiently detect DNA damage in highly dilute samples (0.01 ng microL(-1)), which is beyond the capability of traditional techniques. AFM imaging can uniquely supplement traditional techniques for sensitive measurements of damage to DNA by ionizing radiation.
Ke
Changhong
C
Center for Biologically Inspired Materials and Material Systems and Department of Mechanical, Engineering and Materials Science, Duke University, Durham, NC 27708, USA.
Jiang
Yong
Y
Mieczkowski
Piotr A
PA
Muramoto
Garrett G
GG
Chute
John P
JP
Marszalek
Piotr E
PE
D004247:DNA / Q000302:isolation & purification / Q000528:radiation effects* / Q000648:ultrastructure*; D053904:DNA Breaks, Single-Stranded / Q000528:radiation effects; D004249:DNA Damage; D004278:DNA, Superhelical / Q000302:isolation & purification / Q000528:radiation effects / Q000648:ultrastructure; D004307:Dose-Response Relationship, Radiation; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005720:Gamma Rays / Q000009:adverse effects; D018625:Microscopy, Atomic Force / Q000379:methods*; D036103:Nanotechnology; D010957:Plasmids / Q000302:isolation & purification / Q000528:radiation effects / Q000648:ultrastructure
D004278:DNA, Superhelical; D004247:DNA