Biological dose estimation of UVA laser microirradiation utilizing charged particle-induced protein foci

J. Splinter, B. Jakob, M. Lang, K. Yano, J. Engelhardt, S. W. Hell, D. J. Chen, M. Durante, G. Taucher-Scholz

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The induction of localized DNA damage within a discrete nuclear volume is an important tool in DNA repair studies. Both charged particle irradiation and laser microirradiation (LMI) systems allow for such a localized damage induction, but the results obtained are difficult to compare, as the delivered laser dose cannot be measured directly. Therefore, we revisited the idea of a biological dosimetry based on the microscopic evaluation of irradiation-induced Replication Protein A (RPA) foci numbers. Considering that local dose deposition is characteristic for both LMI and charged particles, we took advantage of the defined dosimetry of particle irradiation to estimate the locally applied laser dose equivalent. Within the irradiated nuclear sub-volumes, the doses were in the range of several hundreds of Gray. However, local dose estimation is limited by the saturation of the RPA foci numbers with increasing particle doses. Even high-resolution 4Pi microscopy did not abrogate saturation as it was not able to resolve single lesions within individual RPA foci. Nevertheless, 4Pi microscopy revealed multiple and distinct 53BP1- and γH2AX-stained substructures within the lesion flanking chromatin domains. Monitoring the local recruitment of the telomere repeat-binding factors TRF1 and TRF2 showed that both proteins accumulated at damage sites after UVA-LMI but not after densely ionizing charged particle irradiation. Hence, our results indicate that the local dose delivered by UVA-LMI is extremely high and cannot be accurately translated into an equivalent ionizing radiation dose, despite the sophisticated techniques used in this study.

Original languageEnglish (US)
Pages (from-to)289-297
Number of pages9
JournalMutagenesis
Volume25
Issue number3
DOIs
StatePublished - 2010

Fingerprint

Charged particles
Lasers
Replication Protein A
Dosimetry
Irradiation
Proteins
Microscopy
Microscopic examination
Telomere-Binding Proteins
DNA
Ionizing radiation
Optical resolving power
Ionizing Radiation
DNA Repair
DNA Damage
Chromatin
Repair
Monitoring

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Splinter, J., Jakob, B., Lang, M., Yano, K., Engelhardt, J., Hell, S. W., ... Taucher-Scholz, G. (2010). Biological dose estimation of UVA laser microirradiation utilizing charged particle-induced protein foci. Mutagenesis, 25(3), 289-297. https://doi.org/10.1093/mutage/geq005

Biological dose estimation of UVA laser microirradiation utilizing charged particle-induced protein foci. / Splinter, J.; Jakob, B.; Lang, M.; Yano, K.; Engelhardt, J.; Hell, S. W.; Chen, D. J.; Durante, M.; Taucher-Scholz, G.

In: Mutagenesis, Vol. 25, No. 3, 2010, p. 289-297.

Research output: Contribution to journalArticle

Splinter, J, Jakob, B, Lang, M, Yano, K, Engelhardt, J, Hell, SW, Chen, DJ, Durante, M & Taucher-Scholz, G 2010, 'Biological dose estimation of UVA laser microirradiation utilizing charged particle-induced protein foci', Mutagenesis, vol. 25, no. 3, pp. 289-297. https://doi.org/10.1093/mutage/geq005
Splinter, J. ; Jakob, B. ; Lang, M. ; Yano, K. ; Engelhardt, J. ; Hell, S. W. ; Chen, D. J. ; Durante, M. ; Taucher-Scholz, G. / Biological dose estimation of UVA laser microirradiation utilizing charged particle-induced protein foci. In: Mutagenesis. 2010 ; Vol. 25, No. 3. pp. 289-297.
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