Highly Sensitive Automated Method for DNA Damage Assessment: Gamma-H2AX Foci Counting and Cell Cycle Sorting
Abstract
:1. Introduction
2. Results and Discussion
2.1. Improved Automated Scoring Classifies Cells According to the Cell Cycle Phase
2.1.1. Characteristic γH2AX Staining Patterns of S- and M-Phase Cells
2.1.2. CENP-F to Identify G2 Cells and Define the Nuclear Area Range of Cells in G2
2.1.3. Pericentrin: The Co-Staining Marker of Choice
2.2. γH2AX Automatic Foci Counting
2.2.1. Foci Analysis
2.2.2. γH2AX Signal Saturation at High Radiation Doses in Non-Sorted Cells
2.2.3. Two Mammography Shots Induce a Significant and Detectable Increase in the Number of Foci
2.2.4. Manual vs. Automatic γH2AX Foci Scoring: A Reliable Automated Approach
3. Discussion
4. Experimental Section
4.1. Cell Cultures
4.2. Irradiation
4.3. Immunostaining
4.4. γH2AX Immunostaining Combined with a BrdU Assay
4.5. Automated Microscopy and Analysis
5. Conclusions
Acknowledgments
Conflict of Interest
References
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Hernández, L.; Terradas, M.; Martín, M.; Tusell, L.; Genescà, A. Highly Sensitive Automated Method for DNA Damage Assessment: Gamma-H2AX Foci Counting and Cell Cycle Sorting. Int. J. Mol. Sci. 2013, 14, 15810-15826. https://doi.org/10.3390/ijms140815810
Hernández L, Terradas M, Martín M, Tusell L, Genescà A. Highly Sensitive Automated Method for DNA Damage Assessment: Gamma-H2AX Foci Counting and Cell Cycle Sorting. International Journal of Molecular Sciences. 2013; 14(8):15810-15826. https://doi.org/10.3390/ijms140815810
Chicago/Turabian StyleHernández, Laia, Mariona Terradas, Marta Martín, Laura Tusell, and Anna Genescà. 2013. "Highly Sensitive Automated Method for DNA Damage Assessment: Gamma-H2AX Foci Counting and Cell Cycle Sorting" International Journal of Molecular Sciences 14, no. 8: 15810-15826. https://doi.org/10.3390/ijms140815810