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Immunostaining for DNA Modifications: Computational Analysis of Confocal Images
JoVE Journal
Genetics
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JoVE Journal Genetics
Immunostaining for DNA Modifications: Computational Analysis of Confocal Images

Immunostaining for DNA Modifications: Computational Analysis of Confocal Images

DOI:
10.3791/56318-v

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09:42 min

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September 07, 2017

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, , , , , , , , ,
1Division of Cancer and Stem Cells, School of Medicine, Centre for Biomolecular Sciences,University of Nottingham, 2School of Life Sciences Imaging (SLIM), School of Life Sciences,University of Nottingham, 3Children’s Brain Tumour Research Centre, School of Medicine, QMC,University of Nottingham

Chapters

  • 00:05Title
  • 00:58Making 2.5D Intensity Plots
  • 03:07Intensity Profiling
  • 04:40Co-localization Analysis
  • 06:43Results: Nuclear Distribution of 5hmC and 5caC Differentiating Cells (Hepatic, Daoy medulloblastoma, BXD-1425EPN Ependymoma, hiPSCs)
  • 08:30Conclusion

Summary

Automatic Translation

Automatic Translation

Newly discovered oxidized forms of 5-methylcytosine (oxi-mCs), 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) may represent distinct DNA modifications with unique functional roles. Here a semi-quantitative workflow for visualization of oxi-mCs' spatial distribution, signal intensity profiling and colocalization is described.

Tags

DNA ModificationsImmunostainingConfocal MicroscopyComputational AnalysisEpigeneticsNuclear LocalizationSignal Intensity2.5D VisualizationImage ProcessingZen Blue Software

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