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Advanced Confocal Microscopy Techniques to Study Protein-protein Interactions and Kinetics at DNA Lesions
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Advanced Confocal Microscopy Techniques to Study Protein-protein Interactions and Kinetics at DNA Lesions

Advanced Confocal Microscopy Techniques to Study Protein-protein Interactions and Kinetics at DNA Lesions

DOI:
10.3791/55999-v

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12:43 min

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November 12, 2017

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, , , ,
1Institute of Biophysics of the Czech Academy of Sciences

Chapters

  • 00:05Title
  • 00:44Cultivation of HeLa-derived Cell Lines
  • 01:48Cell Transfection
  • 03:21Induction of Local DNA Lesions and Confocal Microscopy
  • 04:47Fluorescence Lifetime Image Microscopy
  • 06:24FRET Efficiency Using the FLIM-FRET Script
  • 07:48FRAP Analysis
  • 08:50Results: Analysis of Recruitment of mCherry-tagged PCNA and -53BP1 at DNA Lesions in HeLa Cells
  • 11:00Conclusion

Summary

Automatic Translation

Automatic Translation

Laser microirradiation is a useful tool for studies of DNA repair in living cells. A methodological approach for the use of UVA lasers to induce various DNA lesions is shown. We have optimized a method for local microirradiation that maintains the normal cell cycle; thus, irradiated cells proceed through mitosis.

Tags

Keywords: Confocal MicroscopyProtein-protein InteractionsDNA LesionsDNA RepairLive Cell ImagingFRAPFLIMFLIM-FRETLocal MicroirradiationFucci Cell Cycle SystemHeLa CellsDNA Damage Response

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