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Imaging Polarization in Budding Yeast.

Allison W McClure1, Chi-Fang Wu1, Sam A Johnson2

  • 1Department of Pharmacology and Cancer Biology, Duke University School of Medicine, C359 LSRC, Box 3813, Durham, NC, 27710, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 9, 2016
PubMed
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We developed live-cell imaging techniques to observe yeast cell polarity establishment. This revealed novel dynamic behaviors like multiple polarity clusters and mobile polarity sites during this crucial cellular process.

Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biophysics

Background:

  • Yeast cell polarity establishment is a fundamental process for cell growth and division.
  • Observing rapid, rare events like polarity establishment in live cells presents significant imaging challenges.
  • Existing live-cell imaging methods often face trade-offs between resolution, duration, and phototoxicity.

Purpose of the Study:

  • To develop and apply advanced live-cell imaging methods for detailed observation of yeast polarity establishment.
  • To overcome limitations of phototoxicity and spatiotemporal resolution in imaging fast cellular events.
  • To uncover previously unobserved dynamic behaviors during yeast cell polarization.

Main Methods:

  • Development of novel live-cell imaging protocols for yeast.
Keywords:
Cdc42Cell polarityGFPImagingMicroscopyPolarity establishmentYeast

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  • Cell synchronization strategies to enhance photodamage resistance.
  • High-resolution microscopy to capture rapid cellular dynamics.
  • Quantitative analysis of polarity factor clustering and localization.

    • Main Results:

    • Successful live-cell imaging of yeast polarity establishment with reduced phototoxicity.
    • Discovery of transient intermediate states with multiple polarity clusters.
    • Observation of oscillatory clustering of polarity factors.
    • Identification of mobile, "wandering" polarity sites during the process.

    Conclusions:

    • Advanced live-cell imaging enables the study of rapid cellular events like yeast polarity.
    • Yeast cell polarization is a more dynamic process than previously understood, involving novel intermediates and behaviors.
    • The developed methods provide a powerful tool for future investigations into cell polarity and other fast biological phenomena.