High resolution CryoFESEM of microbial surfaces

Stanley Erlandsen, Ming Lei, Ines Martin-Lacave, Gary M Dunny, Carol L Wells

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

The outer surfaces of three microorganisms, Giardia lamblia, Enterococcus faecalis, and Proteus mirabilis, were investigated by cryo-immobilization followed by sublimation of extracellular ice and cryocoating with either Pt alone or Pt plus carbon. Cryocoated samples were examined at -125°C in either an in-lens field emission SEM or a below-the-lens field emission SEM. Cryocoating with Pt alone was sufficient for low magnification observation, but attempts to do high-resolution imaging resulted in radiolysis and cracking of the specimen surface. Double coating with Pt and carbon, in combination with high resolution backscatter electron detectors, enabled high-resolution imaging of the glycocalyx of bacteria, revealing a sponge-like network over the surface. High resolution examination of bacterial flagella also revealed a periodic substructure. Common artifacts included radiolysis leading to "cracking" of the surface, and insufficient deposition of Pt resulting in the absence of detectable surface topography.

Original languageEnglish (US)
Pages (from-to)273-278
Number of pages6
JournalMicroscopy and Microanalysis
Volume9
Issue number4
DOIs
StatePublished - Aug 1 2003

Keywords

  • Cryo-immobilization
  • CryoFESEM
  • Enterococcus faecalis
  • Extracellular topography
  • Field emission SEM
  • Giardia lamblia
  • Glycocalyx
  • Proteus mirabilis

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