Bringing electrons and microarray technology together.
Low-energy secondary electrons are the most abundant radiolysis species which are thought to be able to attach to and damage DNA via formation and decay of localized molecular resonances involving DNA components. In this study, we analyze the consequences of low-energy electron impact on the ability of DNA to hybridize (i.e., to form the duplex). Specifically, single-stranded thymine DNA oligomers tethered to a gold surface are irradiated with very low-energy electrons (E = 3 eV, which is below the 7.5 eV ionization threshold of DNA) and subsequently exposed to a dye-marked complementary strand to quantify by a fluorescence method the electron induced damage. The damage to (dT)25 oligomers is detected at quite low electron doses with only about 300 electrons per oligomer being sufficient to completely preclude its hybridization. In the microarray format, the method can be used for a rapid screening of the sequence dependence of the DNA-electron interaction. We also show for the first time that the DNA reactions at surfaces can be imaged by secondary electron (SE) emission with both high analytical and spatial sensitivity. The SE micrographs indicate that strand breaks induced by the electrons play a significant role in the reaction mechanism.
Solomun
T
T
Free University Berlin, Institute of Chemistry and Biochemistry, Physical and Theoretical Chemistry, Takustrasse 3, D-14195 Berlin, Germany. solomun@chemie.fu-berlin.de
Sturm
H
H
D004247:DNA / Q000737:chemistry* / Q000528:radiation effects*; D004249:DNA Damage; D004583:Electrons; D005453:Fluorescence; D006046:Gold / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012680:Sensitivity and Specificity; D013499:Surface Properties
D006046:Gold; D004247:DNA