Journal
ANTIBIOTICS-BASEL
Volume 10, Issue 3, Pages -Publisher
MDPI
DOI: 10.3390/antibiotics10030307
Keywords
transmission electron microscopy; bacterial cell wall; bacterial membrane; Gram-negative bacteria; colistin; drug interaction; Tokuyasu cryo-ultramicrotomy
Categories
Funding
- Australian Research Council (ARC) [LP110200770]
- Neoculi Pty Ltd.
- University of South Australia
- Australian Research Council [LP110200770] Funding Source: Australian Research Council
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Through comparing different TEM methods, this study found that ultrathin cryo-sections are superior in visualizing the membrane ultrastructure of Escherichia coli and Pseudomonas aeruginosa. Therefore, ultrathin cryo-sections can be used to better observe and understand the interaction mechanisms of drugs on the bacterial cell membrane.
In this study, we optimized and compared different transmission electron microscopy (TEM) methods to visualize changes to Gram-negative bacterial morphology induced by treatment with a robenidine analogue (NCL195) and colistin combination. Aldehyde-fixed bacterial cells (untreated, treated with colistin or NCL195 + colistin) were prepared using conventional TEM methods and compared with ultrathin Tokuyasu cryo-sections. The results of this study indicate superiority of ultrathin cryo-sections in visualizing the membrane ultrastructure of Escherichia coli and Pseudomonas aeruginosa, with a clear delineation of the outer and inner membrane as well as the peptidoglycan layer. We suggest that the use of ultrathin cryo-sectioning can be used to better visualize and understand drug interaction mechanisms on the bacterial cell membrane.
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