Effect of fixation‐resin combinations and ruthenium red on elucidating outer envelope structure and surface morphology of two methanotrophic bacteria
作者:
Theresa A. Fassel,
Marilyn J. Schaller,
Mary E. Lidstrom,
Charles C. Remsen,
期刊:
Journal of Electron Microscopy Technique
(WILEY Available online 1990)
卷期:
Volume 14,
issue 1
页码: 52-62
ISSN:0741-0581
年代: 1990
DOI:10.1002/jemt.1060140109
出版商: Wiley Subscription Services, Inc., A Wiley Company
关键词: Cell wall;LR White;Spurr
数据来源: WILEY
摘要:
AbstractWe 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 Spurrembedments with the following fixations: glutaraldehyde/OsO4, two glutaraldehyde‐paraformal dehyde, and two different en bloc ruthenium red procedures, one utilizing 0sO4and the other with glutaraldehyde/OsO4in 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 ruthenium red) 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 “spikelike” 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 techniques were not seen in SEM. Presence or absence of particular envelope structures in these methanotrophs is dependent on the combination of fixatives and/or resins employed and is species‐specific. The chemical preparation methods used resulted in enhanced understanding of the structure and composition of
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