Electron microscopy of frozen-hydrated bacteria

Details

Serval ID
serval:BIB_19DBA4D565CB
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Electron microscopy of frozen-hydrated bacteria
Journal
Journal of Bacteriology
Author(s)
Dubochet  J., McDowall  A. W., Menge  B., Schmid  E. N., Lickfeld  K. G.
ISSN
0021-9193 (Print)
Publication state
Published
Issued date
07/1983
Volume
155
Number
1
Pages
381-90
Notes
Journal Article --- Old month value: Jul
Abstract
Amorphous, unstained, frozen-hydrated sections of bacteria provide a faithful high-resolution image of procaryotic cells. Conventional preparation artifacts due to fixation, staining, and dehydration are nonexistent. Freezing damage is avoided by using glucose as a cryoprotectant. Cutting damage on frozen material is severe, but sectioning artifacts, being always related to the cutting direction, can be systematically recognized and thus taken into consideration. Geometry and density distribution of the bacterial envelope can be resolved to about 3 nm. The following main features have been observed. In Escherichia coli the inner and outer membranes have an approximately uniform density profile. The distance between the two membranes is constant, ca. 33 nm. In Staphylococcus aureus the cell wall is ca. 40 nm wide. It is bordered on the cytoplasmic side by an asymmetric 5.5-nm-wide bilayer. The bacterial nucleoid, clearly visible with conventional preparation methods, appears in exponentially growing bacteria as an ill-defined central region with approximately the same density as the rest of the cytoplasm. It becomes more clearly visible when bacteria are in the stationary phase, plasmolysed, fixed, or stained. We confirm that "mesosomes," hitherto quite often considered to be essential organelles in all procaryotes, are artifacts. They appear in large numbers during osmium fixation.
Keywords
Bacillus subtilis/ultrastructure Bacteria/*ultrastructure Escherichia coli/ultrastructure Freezing Klebsiella pneumoniae/ultrastructure Microscopy, Electron/methods Staphylococcus aureus/ultrastructure
Pubmed
Web of science
Create date
24/01/2008 10:25
Last modification date
20/08/2019 12:50
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