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Related Experiment Videos

Method for immobilizing microbial cells on gel surface for dynamic AFM studies

M Gad1, A Ikai

  • 1Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.

Biophysical Journal
|December 1, 1995
PubMed
Summary

Atomic force microscopy successfully imaged Saccharomyces cerevisiae cell growth and budding over 6-7 hours. Immobilized yeast cells showed growth, indicating the atomic force microscope method effectively captured the dynamic cell processes without significant interference.

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Area of Science:

  • Microbiology
  • Cell Biology
  • Biophysics

Background:

  • Understanding yeast cell dynamics is crucial for various biotechnological applications.
  • Previous imaging techniques faced limitations in resolving live cell processes with high fidelity.

Purpose of the Study:

  • To image the dynamic processes of cell growth and budding in Saccharomyces cerevisiae using atomic force microscopy (AFM).
  • To assess the reliability of AFM for long-term live cell imaging without significant interference.

Main Methods:

  • Yeast cells (Saccharomyces cerevisiae) were gently immobilized on 3% agar and submerged in culture medium.
  • Atomic force microscopy (AFM) was employed to image the immobilized cells over a period of 6-7 hours.
  • Control experiments using chemically fixed cells were performed to evaluate potential artifacts.

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Main Results:

  • Live yeast cells exhibited observable growth, including increments in height and lateral enlargement.
  • A growing daughter cell attached to its mother cell was successfully imaged, demonstrating the visualization of budding.
  • Chemically fixed cells showed minimal changes, indicating that agar dissolution or tip-scraping did not significantly affect imaging of live cells.

Conclusions:

  • Atomic force microscopy provides a viable method for imaging the dynamic growth and budding processes of Saccharomyces cerevisiae.
  • The gentle immobilization technique on agar allows for extended observation of live yeast cell morphology and development.
  • AFM imaging is reliable for studying live cell processes, offering insights into cell division and morphology changes over time.