Nanoscale focused ion beam tomography of single bacterial cells for assessment of antibiotic effects

Liu, Boyin, Yu, Heidi H., Ng, Tuck Wah, Paterson, David L., Velkov, Tony, Li, Jian and Fu, Jing (2014) Nanoscale focused ion beam tomography of single bacterial cells for assessment of antibiotic effects. Microscopy and Microanalysis, 20 2: 537-547. doi:10.1017/S1431927614000026

Author Liu, Boyin
Yu, Heidi H.
Ng, Tuck Wah
Paterson, David L.
Velkov, Tony
Li, Jian
Fu, Jing
Title Nanoscale focused ion beam tomography of single bacterial cells for assessment of antibiotic effects
Journal name Microscopy and Microanalysis   Check publisher's open access policy
ISSN 1431-9276
Publication date 2014-04-01
Year available 2014
Sub-type Article (original research)
DOI 10.1017/S1431927614000026
Open Access Status DOI
Volume 20
Issue 2
Start page 537
End page 547
Total pages 11
Place of publication Cambridge, United Kingdom
Publisher Cambridge University Press
Language eng
Subject 3105 Instrumentation
Abstract Antibiotic resistance is a major risk to human health, and to provide valuable insights into mechanisms of resistance, innovative methods are needed to examine the cellular responses to antibiotic treatment. Focused ion beam tomography is proposed to image and assess the detailed three-dimensional (3D) ultrastructure of single bacterial cells. By iteratively removing slices of thickness in the order of 10 nm, high magnification 2D images can be acquired by scanning electron microscopy at single-digit nanometer resolution. In this study, Klebsiella pneumoniae was treated with polymyxin B, and 3D models of both cell envelope and cytoplasm regions containing the nucleoid and ribosomes were reconstructed. The 3D volume containing the nucleoid and ribosomes was significantly smaller, and the cell length along the longitudinal axis was extended by 40% in the treated cells, implying stress responses to the drug treatment. More than a 200% increase in protrusions per unit surface area on the cell envelope was observed in the curvature analysis after treatment. Experiments by conventional transmission electron microscopy and atomic force microscopy were also performed, followed by comparison and discussions. In conclusion, the proposed 3D imaging method and associated analysis provide a unique tool for the assessment of antibiotic effects on multidrug-resistant bacteria at nanometer resolution.
Keyword Tomography
Antibiotic resistance
Curvature analysis
Electron microscopy
Focused ion beam
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: UQ Centre for Clinical Research Publications
Official 2015 Collection
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Citation counts: TR Web of Science Citation Count  Cited 4 times in Thomson Reuters Web of Science Article | Citations
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