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Sample preparation for electron microscopy of internal cell structure.

G H Haggis1

  • 1Research Branch, Agriculture Canada, Ottawa.

Microscopy Research and Technique
|July 1, 1992
PubMed
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High-resolution scanning electron microscopy and rapid-freeze deep-etch techniques reveal internal cell structures. Permeabilizing cells before imaging is crucial for detailed 3D ultrastructure studies.

Area of Science:

  • Cell Biology
  • Microscopy Techniques

Background:

  • Examining internal cell structure is vital for understanding cellular functions.
  • Traditional electron microscopy methods face challenges with complex cellular matrices.

Purpose of the Study:

  • To review and compare methods for high-resolution imaging of internal cell ultrastructure.
  • To identify optimal techniques for visualizing functional cellular processes.

Main Methods:

  • Comparison of high-resolution scanning electron microscopy (SEM) with rapid-freeze deep-etch replica transmission electron microscopy (TEM).
  • Evaluation of freeze-fracture techniques for ultrastructure studies.
  • Assessment of cell permeabilization and fixation protocols.

Main Results:

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  • Rapid freezing alone is often insufficient for deep 3D views due to cellular composition.
  • Cell permeabilization, followed by fixation, is essential for effective imaging.
  • Both deep-etch replication and high-resolution SEM can reveal detailed structures after preparation.

Conclusions:

  • Permeabilizing cultured cells preserves structure and function for detailed ultrastructural analysis.
  • Combined permeabilization and fixation enable advanced imaging by SEM or deep-etch TEM.
  • These methods provide deep views of cellular components, such as chromosomes and Golgi transport.