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High Resolution CryoFESEM of Microbial Surfaces

Published online by Cambridge University Press:  01 August 2003

Stanley Erlandsen
Affiliation:
Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, MN 55455
Ming Lei
Affiliation:
Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, MN 55455
Ines Martin-Lacave
Affiliation:
Departmento Citología e Histología Normal y Patológica, Facultad de Medicina, Universidad de Sevilla, Sevilla, Spain
Gary Dunny
Affiliation:
Department of Microbiology, University of Minnesota Medical School, Minneapolis, MN 55455
Carol Wells
Affiliation:
Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, MN 55455
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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.

Type
High Resolution Cryo-SEM
Copyright
© 2003 Microscopy Society of America

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