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Field-emission scanning electron microscopy of the internal cellular organization of fungi

W H Müller1, A C van Aelst, B M Humbel

  • 1Department of Molecular Cell Biology, EMSA, Utrecht University, The Netherlands. W.H.Muller@bio.uu.nl

Scanning
|October 7, 2000
PubMed

Insights

Scanning electron microscopy (SEM) offers a new way to view fungal cell structures. This method reveals detailed intracellular ultrastructure, including organelles and septal pore caps, in fungi.

Area of Science:

  • Mycology
  • Cell Biology
  • Microscopy

Background:

  • Transmission electron microscopy (TEM) of sectioned fungal material is a standard method for studying hyphal cellular organization.
  • Observing intact fungal cells offers an alternative perspective to sectioned material.

Purpose of the Study:

  • To develop and validate a scanning electron microscopy (SEM) method for visualizing the intracellular ultrastructure of fungal hyphae.
  • To provide a detailed internal view of fungal cellular organization as an alternative to traditional TEM methods.

Main Methods:

  • Fungal colonies were chemically fixed (glutaraldehyde, osmium tetroxide) and treated with dimethyl sulfoxide.
  • Samples underwent freeze-fracturing, maceration in osmium tetroxide to remove the cytoplasmic matrix, and freeze-substitution dehydration.
  • Final processing included critical point drying, mounting, sputter coating, and imaging with field-emission SEM.

Main Results:

  • The SEM method successfully produced clear images of intracellular fungal structures.
  • Visualized organelles included mitochondria (elongated and spherical), the nucleus, and intravacuolar structures.
  • Detailed views of tubular and plate-like endoplasmic reticulum and various septal pore caps were obtained.

Conclusions:

  • This SEM procedure is a powerful approach for studying the intracellular ultrastructure of fungi.
  • It provides high-resolution imaging of organelles and cellular components within intact hyphae.
  • This technique offers a valuable alternative for fungal cell biology research.

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