title: Visualization of heavy ion tracks by labeling 3'-OH termini of induced DNA strand breaks.
abstract: African green monkey kidney cells, CV-1, were irradiated with Carbon ions (LET: 735 keV/µm Argon ions (LET: 3,000 keV/µm) to visualize ion tracks through the cell nucleus by labeling the 3'-OH termini result of DNA strand breaks. The 3'-OH termini of DNA were labeled with BrdU-triphosphate catalyzed by TdT. This method of TUNEL (TdT-mediated dUTP Nick End labeling) is based on the specific binding of TdT to 3'-OH termini of DNA. Subsequent immuno-fluorescent staining with the primary monoclonal antibody against BrdU, followed by a secondary antibody of Alexa Fluor 488, was performed to visualize the BrdU labeled DNA termini. Images of the cell nuclei were acquired by confocal laser microscopy. When cell monolayers were irradiated perpendicularly with argon ions, induced DSBs in cell nuclei were identifiable as fluorescent spots. In another irradiation setup, when cells were irradiated at a small angle with incident argon ions, DNA strand breaks were detected as fluorescent stripes across the cell nucleus. These results demonstrate the induction of 3'-OH termini at sites of DNA strand breaks along Argon ion tracks.
authors.affiliation: Department of Technical Support and Development, Fundamental Technology Center, National Institute of Radiological Sciences. tkonishi@nirs.go.jp
authors.forename: Teruaki
authors.initials: T
authors.lastname: Konishi
authors.forename: Akihiro
authors.initials: A
authors.lastname: Takeyasu
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Natsume
authors.forename: Yoshiya
authors.initials: Y
authors.lastname: Furusawa
authors.forename: Kotaro
authors.initials: K
authors.lastname: Hieda
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001128:Argon; D002244:Carbon / Q000737:chemistry; D002467:Cell Nucleus / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004249:DNA Damage; D005456:Fluorescent Dyes / Q000494:pharmacology; D005720:Gamma Rays; D020287:In Situ Nick-End Labeling; D007477:Ions; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D013388:Succinimides / Q000494:pharmacology
chemical_list: C492694:Alexa Fluor 488 carboxylic acid succinimidyl ester; D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D007477:Ions; D013388:Succinimides; D001128:Argon; D002244:Carbon