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Temporal Tracking of Cell Cycle Progression Using Flow Cytometry without the Need for Synchronization
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Temporal Tracking of Cell Cycle Progression Using Flow Cytometry without the Need for Synchronization.

Robert Welschinger1, Linda J Bendall2

  • 1Centre for Cancer Research, Westmead Millennium Institute for Medical Research and University of Sydney.

Journal of Visualized Experiments : Jove
|September 2, 2015
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Summary

This study presents a novel method for tracking cells through the cell cycle using bromodeoxyuridine (BrdU) incorporation, avoiding toxic synchronization. This technique allows for precise determination of cell cycle phase durations and protein expression without harming cell populations.

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Area of Science:

  • Cell Biology
  • Molecular Biology

Background:

  • Cell synchronization is a common but often toxic method for cell cycle analysis.
  • Some cell types are sensitive to synchronization blocks, leading to abnormal cycling or cell death.

Purpose of the Study:

  • To describe a method for tracking cells through the cell cycle without synchronization.
  • To enable determination of cell cycle phase durations and protein expression in individual cells.

Main Methods:

  • Utilizing bromodeoxyuridine (BrdU) uptake to label cells during the S phase of DNA synthesis.
  • Tracking BrdU-labeled cells through subsequent cell cycle stages.
  • Correlating protein expression with cell cycle progression.

Main Results:

  • Successfully tracked cells through the cell cycle without synchronization.
  • Determined cell cycle phase durations for individual cells.
  • Enabled assessment of protein expression and cellular functions at different cell cycle stages.

Conclusions:

  • The BrdU-based method provides a non-toxic alternative to cell synchronization for cell cycle analysis.
  • This protocol allows for precise measurement of cell cycle kinetics and assessment of cellular responses.