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Quantitative analysis of DNA-damage response factors after sequential ion microirradiation

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Abstract

Several proteins are known to form foci at DNA sites damaged by ionizing radiation. We study DNA damage response by immunofluorescence microscopy after microirradiation of cells with energetic ions. By using microirradiation, it is possible to irradiate different regions on a single dish at different time-points and to differentiate between cells irradiated earlier and later. This allows to directly compare immunofluorescence intensities in both subsets of cells with little systematic error because both subsets are cultivated and stained under identical conditions. In addition, by using irradiation patterns such as crossing lines, it is possible to irradiate individual cells twice and to differentiate between immunofluorescence signals resulting from the cellular response to the earlier and to the later irradiation event. Here, we describe the quantitative evaluation of immunofluorescence intensities after sequential irradiation.

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Acknowledgments

We thank the staff of the Maier-Leibnitz-Laboratorium for operating the accelerator and F. Eckardt-Schupp, Helmholtz Zentrum München, for providing access to the α-irradiation facility. This work was supported by grants of the European Science Foundation (ESF) under the EUROCORES Programme EuroDYNA (G.D., T.C., A.A.F.), the DFG Cluster of Excellence Munich Centre for Advanced Photonics (G.D., A.A.F.), the Bundesamt für Strahlenschutz (A.A.F.) and the Bundesministerium für Bildung und Forschung (A.A.F.).

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Correspondence to Christoph Greubel.

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Greubel, C., Hable, V., Drexler, G.A. et al. Quantitative analysis of DNA-damage response factors after sequential ion microirradiation. Radiat Environ Biophys 47, 415–422 (2008). https://doi.org/10.1007/s00411-008-0181-0

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  • DOI: https://doi.org/10.1007/s00411-008-0181-0

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