PMID: 7938938Jan 1, 1994Paper

Use of freeze-fracturing to observe the ultrastructure of Candida albicans with scanning electron microscopy

Revista latinoamericana de microbiología
S A PortilloB Araujo

Abstract

The development of the techniques of sample preparation for the observation of ultrastructure with scanning electron microscopy (SEM) include, among others, the method O-D-O reported by Tanaka and Naguro in 1981, based on the initial fixing of the specimen (eukaryotic tissue) in 1% OsO4; treatment with dimethyl sulphoxide (DMSO) as a cryoprotectant, followed by cryofracture, removal of cytoplasmatic matrix by psot-fixing with 1% OsO4 and a further conductive stain. In this paper we report the application of the method O-D-O in a culture of Candida albicans modifying the pre-fixing carried out overnight and excepting the osmic maceration, in order to observe the cell ultrastructure of this eukaryotic microorganism with SEM at a normal resolution. Through the tridimensional images obtained, in which we can observe the fracture of Candida albicans without cytoplasmatic matrix but with details of a normal and deformed cell wall, as well as cells conserving their cytoplasmatic matrix where globose and filamentous structures become evident and suggest a pattern of organization and cells with a very clear character of what we believe is the nuclear zone of the microorganism, we night conclude that our method is suitable for eukaryotic...Continue Reading

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