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Related Experiment Videos

DNA labeling in living cells.

Robert M Martin1, Heinrich Leonhardt, M Cristina Cardoso

  • 1Max Delbrück Center for Molecular Medicine, Franz-Volhard-Klinik, Berlin, Germany.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|August 6, 2005
PubMed
Summary

The DNA dye DRAQ5 is ideal for live cell imaging, offering low photobleaching and excellent chromatin visualization. It is compatible with other fluorescent proteins, making it a versatile tool for nuclear studies.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Molecular Imaging

Background:

  • Live cell fluorescence microscopy requires effective visualization of the nucleus and chromatin.
  • Determining nuclear morphology and the localization of nuclear compartments is crucial in cell biology.

Purpose of the Study:

  • To evaluate the suitability of five DNA dyes for live cell imaging.
  • To compare DNA dyes with fluorescently labeled histones for chromatin marking.

Main Methods:

  • Tested TOPRO-3, TOTO-3, propidium iodide, Hoechst 33258, and DRAQ5 in live cells.
  • Utilized widefield epifluorescence and confocal laser scanning microscopy with continuous imaging and photobleaching.
  • Compared DNA staining with histone-GFP fusion proteins.

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Main Results:

  • Only Hoechst 33258 and DRAQ5 effectively stained DNA in living cells.
  • DRAQ5 exhibited low photobleaching and comparable chromatin compartment labeling to H2B-GFP.
  • DRAQ5's deep red excitation/emission is compatible with common fluorescent proteins (e.g., C/G/YFP, mRFP).

Conclusions:

  • DRAQ5 is a suitable DNA dye for chromatin visualization in living cells.
  • DRAQ5 can be easily combined with fluorophores emitting in the blue to orange spectrum.