title: Co-visualization of DNA damage and ion traversals in live mammalian cells using a fluorescent nuclear track detector.
abstract: The geometric locations of ion traversals in mammalian cells constitute important information in the study of heavy ion-induced biological effect. Single ion traversal through a cellular nucleus produces complex and massive DNA damage at a nanometer level, leading to cell inactivation, mutations and transformation. We present a novel approach that uses a fluorescent nuclear track detector (FNTD) for the simultaneous detection of the geometrical images of ion traversals and DNA damage in single cells using confocal microscopy. HT1080 or HT1080-53BP1-GFP cells were cultured on the surface of a FNTD and exposed to 5.1-MeV/n neon ions. The positions of the ion traversals were obtained as fluorescent images of a FNTD. Localized DNA damage in cells was identified as fluorescent spots of γ-H2AX or 53BP1-GFP. These track images and images of damaged DNA were obtained in a short time using a confocal laser scanning microscope. The geometrical distribution of DNA damage indicated by fluorescent γ-H2AX spots in fixed cells or fluorescent 53BP1-GFP spots in living cells was found to correlate well with the distribution of the ion traversals. This method will be useful for evaluating the number of ion hits on individual cells, not only for micro-beam but also for random-beam experiments.
authors.affiliation: Research, Development and Support Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inageku, Chiba 263-8555, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kodaira
authors.affiliation: Research, Development and Support Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inageku, Chiba 263-8555, Japan tkonishi@nirs.go.jp.
authors.forename: Teruaki
authors.initials: T
authors.lastname: Konishi
authors.affiliation: Research, Development and Support Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inageku, Chiba 263-8555, Japan.
authors.forename: Alisa
authors.initials: A
authors.lastname: Kobayashi
authors.affiliation: Research, Development and Support Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inageku, Chiba 263-8555, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Maeda
authors.affiliation: Division of Agrotechnology and Biosciences, Malaysian Nuclear Agency, Bangi, Kajang 43000, Malaysia.
authors.forename: Tengku Ahbrizal Farizal Tengku
authors.initials: TA
authors.lastname: Ahmad
authors.affiliation: State Key Laboratory of Nuclear Physics and Technology, Peking University, Beijing 100871, PR China.
authors.forename: Gen
authors.initials: G
authors.lastname: Yang
authors.affiliation: Landauer Inc., Crystal Growth Division, Stillwater, OK 74074, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Akselrod
authors.affiliation: Research, Development and Support Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inageku, Chiba 263-8555, Japan.
authors.forename: Yoshiya
authors.initials: Y
authors.lastname: Furusawa
authors.affiliation: Research, Development and Support Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inageku, Chiba 263-8555, Japan.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Uchihori
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics / Q000528:radiation effects; D004249:DNA Damage / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005354:Fibrosarcoma / Q000473:pathology* / Q000503:physiopathology*; D006801:Humans; D007477:Ions; D018499:Linear Energy Transfer / Q000528:radiation effects; D018613:Microscopy, Confocal / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D011829:Radiation Dosage; D011874:Radiometry / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D007477:Ions
keywords: DNA damage; fluorescent nuclear track detector; ion beam tracks; localization