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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Updated: Jun 19, 2026

Preparation and Cryo-FIB micromachining of Saccharomyces cerevisiae for Cryo-Electron Tomography
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Preparation and Cryo-FIB micromachining of Saccharomyces cerevisiae for Cryo-Electron Tomography

Published on: November 20, 2021

FINE STRUCTURE IN FROZEN-ETCHED YEAST CELLS.

H Moor1, K Mühlethaler

  • 1Laboratory of Electron Microscopy, Department of General Botany, Swiss Federal Institute of Technology, Zürich, Switzerland.

The Journal of Cell Biology
|October 30, 2009
PubMed
Summary

Freeze-etching electron microscopy reveals high-fidelity yeast cell structures without killing them. This technique visualizes novel surface details of membranes and organelles, offering insights into cellular organization and artifacts.

Area of Science:

  • Cell Biology
  • Microscopy Techniques

Background:

  • Traditional chemical fixation methods for electron microscopy can introduce artifacts.
  • Investigating cellular structures in a life-like state is crucial for understanding their function.

Purpose of the Study:

  • To develop and apply a non-lethal freeze-fixation technique for high-fidelity electron microscopy of yeast cells (Saccharomyces cerevisiae).
  • To reveal previously unobserved surface structures of yeast cell membranes and organelles.
  • To compare freeze-etching results with conventional fixation methods.

Main Methods:

  • Application of a specialized freeze-fixation technique for yeast cells, preserving cellular viability.
  • Utilizing freeze-etching (a form of freeze-drying) for electron microscopic investigation.

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Cryosectioning Yeast Communities for Examining Fluorescence Patterns
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  • Analysis of both cross-fractured and surface views of cellular components.

    • Main Results:

    • Detailed visualization of yeast cell membranes and organelles, including novel surface structures.
    • Identification of hexagonal particle arrangements in the cytoplasmic membrane potentially involved in glucan fibril production.
    • Observation of age-dependent alterations in nuclear pore distribution.
    • Clear distinction of endoplasmic reticulum and vacuoles.
    • Ribosome-covered membranes of vesicular systems and perforations in the mitochondrial envelope.
    • Characterization of storage granules as concentric lipid layers and detection of a potential Golgi apparatus role in lipid storage.
    • Confirmation of unit membrane structure and identification of glycogen agglomerations in older cells.
    • Insights into artifacts caused by chemical fixation and embedding.

    Conclusions:

    • Freeze-etching provides high-fidelity images of yeast cells, revealing intricate structures and cellular processes.
    • The technique offers superior visualization of membrane-associated particles and organelle details compared to traditional methods.
    • This approach aids in understanding cellular organization, identifying artifacts, and exploring the dynamic nature of cellular components.