RRC ID 1161
Author Yamaguchi M, Ohkusu M, Sameshima M, Kawamoto S.
Title Safe specimen preparation for electron microscopy of pathogenic fungi by freeze-substitution after glutaraldehyde fixation.
Journal Nihon Ishinkin Gakkai Zasshi
Abstract A safe method is described for observing ultrastructure of highly infectious fungi by ultrathin sectioning electron microscopy. The fungal cells were first chemically fixed by glutaraldehyde to kill them. They were then rapidly frozen by propane slush in liquid nitrogen and freeze-substituted in acetone containing 2% osmium tetroxide. This method gave clear cell images with high resolution in a natural state, close to the image obtained by rapidly frozen freeze-substituted specimen of living cells. Although we have demonstrated the utility of this method using Exophiala dermatitidis and Cryptococcus neoformans, it could also be used for observing highly infectious fungi such as Coccidioides immitis.
Volume 46(3)
Pages 187-92
Published 2005-1-1
DOI 10.3314/jjmm.46.187
PMID 16094294
MeSH Freeze Substitution* Fungi / ultrastructure* Glutaral / pharmacology* Microscopy, Electron* Specimen Handling / methods* Tissue Fixation
Resource
Pathogenic microorganisms IFM 4844 IFM 5805 IFM 5844 IFM 41464