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Expansion Microscopy on Saccharomyces cerevisiae.

Artemis G Korovesi1, Leonor Morgado1, Marco Fumasoni1

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We developed an improved expansion microscopy (ExM) protocol for yeast (Saccharomyces cerevisiae) to visualize subcellular structures. This method overcomes challenges posed by the yeast cell wall, enabling clearer imaging of cellular ultrastructure.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Yeast Genetics

Background:

  • Saccharomyces cerevisiae is a key model organism for eukaryotic studies.
  • Its small size and resilient cell wall impede detailed subcellular imaging.
  • Existing expansion microscopy (ExM) protocols struggle with yeast due to cell wall interference.

Purpose of the Study:

  • To develop and detail a modified ExM protocol for S. cerevisiae.
  • To enable high-resolution visualization of yeast intracellular architecture.
  • To achieve isotropic expansion overcoming cell wall limitations.

Main Methods:

  • A modified expansion microscopy (ExM) protocol was developed for S. cerevisiae.
  • Critical steps focus on cell wall digestion to achieve uniform sample expansion.
  • The protocol involves sample fixation, embedding in a swellable gel, and isotropic expansion.

Main Results:

  • The protocol achieves a 4x expansion factor for yeast samples.
  • It enables visualization of subcellular ultrastructure with improved clarity.
  • The method ensures isotropic expansion, crucial for accurate structural analysis.

Conclusions:

  • This modified ExM protocol effectively addresses the challenges of imaging yeast.
  • It provides a valuable tool for studying yeast cell biology at an ultrastructural level.
  • The detailed procedure facilitates wider adoption for high-resolution yeast imaging.