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5-Ethynyl-2′-Deoxyuridine/Phospho-Histone H3 Dual-Labeling Protocol for Cell Cycle Progression Analysis in Drosophila Neural Stem Cells
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5-Ethynyl-2′-Deoxyuridine/Phospho-Histone H3 Dual-Labeling Protocol for Cell Cycle Progression Analysis in Drosophila Neural Stem Cells

5-Ethynyl-2′-Deoxyuridine/Phospho-Histone H3 Dual-Labeling Protocol for Cell Cycle Progression Analysis in Drosophila Neural Stem Cells

DOI:
10.3791/62642-v

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09:34 min

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May 04, 2021

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,
1Cell Biology,University of Bern, 2Department of life sciences and medicine,University of Luxembourg

Chapters

  • 00:04Introduction
  • 00:40Dissection of Third Instar Larval Brains and 5-ethynyl-2 -deoxyuridine (EdU) Incorporation
  • 02:23EdU Detection, DNA Staining, and Mounting
  • 04:29Imaging
  • 05:52Image Analysis
  • 08:06Results: EdU and pH3 Dual-labeling for Cell Cycle Progression Analysis in Drosophila Neural Stem Cells
  • 09:01Conclusion

Summary

Automatic Translation

Automatic Translation

Cell cycle analysis with 5-ethynyl-2'-deoxyuridine (EdU) and phospho-histone H3 (pH3) labeling is a multi-step procedure that may require extensive optimization. Here, we present a detailed protocol that describes all steps for this procedure including image analysis and quantification to distinguish cells in different cell cycle phases.

Tags

Keywords: 5-Ethynyl-2′-DeoxyuridinePhospho-Histone H3Cell CycleNeural Stem CellsDrosophilaImmunostainingImage AcquisitionImage ProcessingMitosisG1S PhaseM PhaseSchneider’s Dissection MediumPBSForcepsDissection MicroscopeEDUFixationPBSTHoechst 33342

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