Comparison of Two Transmission Electron Microscopy Methods to Visualize Drug-Induced Alterations of Gram-Negative Bacterial Morphology.
Hang Thi NguyenLisa A O'DonovanHenrietta VenterCecilia C RussellAdam McCluskeyStephen W PageDarren J TrottAbiodun David OgunniyiPublished in: Antibiotics (Basel, Switzerland) (2021)
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.
Keyphrases
- electron microscopy
- gram negative
- multidrug resistant
- escherichia coli
- pseudomonas aeruginosa
- drug induced
- acinetobacter baumannii
- liver injury
- klebsiella pneumoniae
- drug resistant
- biofilm formation
- cystic fibrosis
- emergency department
- high efficiency
- cell death
- cell cycle arrest
- adverse drug
- cell wall
- replacement therapy
- smoking cessation
- single molecule
- fluorescent probe