Effect of fixation-resin combinations and ruthenium red on elucidating outer envelope structure and surface morphology of two methanotrophic bacteria

Journal of Electron Microscopy Technique
T A FasselC C Remsen

Abstract

We examined the ultrastructure of the cell envelope in Type I, Methylomonas albus (BG8), and Type II, Methylosinus trichosporium (OB3b), methane-oxidizing bacteria by using different fixatives, ruthenium red (RR) combinations and resins. We compared LR White and Spurr embedments with the following fixations: glutaraldehyde/OsO4, two glutaraldehyde-paraformaldehyde, and two different en bloc ruthenium red procedures, one utilizing OsO4 and the other with glutaraldehyde/OsO4 in sequential fixation. These fixations were also studied by scanning electron microscopy (SEM). Unfixed cells prepared by freeze etch were used for comparison. Transmission electron microscopy of BG8 embedded in LR White resin (with or without red0 preserved a layer of cup-like structures that were not seen in Spurr resin-embedded cells unless ruthenium red was used. For OB3b, the second RR method preserved beads and filaments where only "spike-like" structures were seen in all other fixations in both resins. By SEM, all fixations preserved a capsular slime layer of BG8 that was removed from some cells by both RR methods. In all SEM fixations, a bead layer was preserved in OB3b that was enhanced by RR. Filaments seen by freeze-etch and thin-section technique...Continue Reading

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Jan 1, 1992·Electron Microscopy Reviews·L L Graham
Mar 8, 2014·Archives of Microbiology·Isty Adhitya PurwasenaKyuro Sasaki
Nov 11, 1999·Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission·T A Fassel, C E Edmiston
Aug 9, 2007·International Journal of Systematic and Evolutionary Microbiology·Angela S LindnerDavid J Hodson
Apr 1, 1991·Applied and Environmental Microbiology·M L CollinsC C Remsen

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