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Related Experiment Videos

High resolution CryoFESEM of microbial surfaces.

Stanley Erlandsen1, Ming Lei, Ines Martin-Lacave

  • 1Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, MN 55455, USA. erlan001@umn.edu

Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada
|August 7, 2003
PubMed
Summary

Cryocoating with platinum and carbon enabled high-resolution imaging of microbial surfaces, revealing detailed structures like bacterial glycocalyx and flagella. This method overcomes limitations of single platinum coating for detailed surface topography analysis.

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Area of Science:

  • Microbiology
  • Microscopy
  • Surface Science

Background:

  • Investigating microbial surface structures is crucial for understanding their function and interactions.
  • Previous imaging techniques faced limitations in resolving fine surface details of microorganisms.

Purpose of the Study:

  • To optimize cryocoating techniques for high-resolution scanning electron microscopy (SEM) of microbial surfaces.
  • To reveal detailed ultrastructural features of Giardia lamblia, Enterococcus faecalis, and Proteus mirabilis.

Main Methods:

  • Cryo-immobilization of microorganisms followed by ice sublimation.
  • Cryocoating with platinum (Pt) alone or Pt plus carbon.
  • Examination using field emission scanning electron microscopy (FESEM) at -125°C.

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  • Utilizing high-resolution backscatter electron detectors.

    • Main Results:

    • Single Pt coating was adequate for low magnification but led to radiolysis and cracking at high resolution.
    • Double coating with Pt and carbon enabled high-resolution imaging of bacterial glycocalyx, showing a sponge-like network.
    • High-resolution examination revealed periodic substructure in bacterial flagella.
    • Identified common artifacts: radiolysis-induced cracking and insufficient Pt deposition.

    Conclusions:

    • Dual Pt/C cryocoating is superior for high-resolution SEM of microbial surfaces.
    • This technique allows detailed visualization of microbial surface topography and ultrastructure.
    • Optimized cryocoating methods enhance the study of microbial surface features.