Near-atomic three-dimensional mapping for site-specific chemistry of 'superbugs'

Adineh, Vahid R., Marceau, Ross K. W., Velkov, Tony, Li, Jian and Fu, Jing 2016, Near-atomic three-dimensional mapping for site-specific chemistry of 'superbugs', Nano letters, vol. 16, no. 11, pp. 7113-7120, doi: 10.1021/acs.nanolett.6b03409.

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Title Near-atomic three-dimensional mapping for site-specific chemistry of 'superbugs'
Author(s) Adineh, Vahid R.
Marceau, Ross K. W.ORCID iD for Marceau, Ross K. W.
Velkov, Tony
Li, Jian
Fu, Jing
Journal name Nano letters
Volume number 16
Issue number 11
Start page 7113
End page 7120
Total pages 8
Publisher American Chemical Society
Place of publication Washington, D.C.
Publication date 2016
ISSN 1530-6992
Keyword(s) Cell imaging
atom probe tomography
drug resistance
nanoscale chemical mapping
Science & Technology
Physical Sciences
Chemistry, Multidisciplinary
Chemistry, Physical
Nanoscience & Nanotechnology
Materials Science, Multidisciplinary
Physics, Applied
Physics, Condensed Matter
Science & Technology - Other Topics
Materials Science
Summary Emergence of multidrug resistant Gram-negative bacteria has caused a global health crisis and last-line class of antibiotics such as polymyxins are increasingly used. The chemical composition at the cell surface plays a key role in antibiotic resistance. Unlike imaging the cellular ultrastructure with well-developed electron microscopy, the acquisition of a high-resolution chemical map of the bacterial surface still remains a technological challenge. In this study, we developed an atom probe tomography (APT) analysis approach to acquire mass spectra in the pulsed-voltage mode and reconstructed the 3D chemical distribution of atoms and molecules in the subcellular domain at the near-atomic scale. Using focused ion beam (FIB) milling together with micromanipulation, site-specific samples were retrieved from a single cell of Acinetobacter baumannii prepared as needle-shaped tips with end radii less than 60 nm, followed by a nanoscale coating of silver in the order of 10 nm. The significantly elevated conductivity provided by the metallic coating enabled successful and routine field evaporation of the biological material, with all the benefits of pulsed-voltage APT. In parallel with conventional cryo-TEM imaging, our novel approach was applied to investigate polymyxin-susceptible and -resistant strains of A. baumannii after treatment of polymyxin B. Acquired atom probe mass spectra from the cell envelope revealed characteristic fragments of phosphocholine from the polymyxin-susceptible strain, but limited signals from this molecule were detected in the polymyxin-resistant strain. This study promises unprecedented capacity for 3D nanoscale imaging and chemical mapping of bacterial cells at the ultimate 3D spatial resolution using APT.
Language eng
DOI 10.1021/acs.nanolett.6b03409
Field of Research 110199 Medical Biochemistry and Metabolomics not elsewhere classified
MD Multidisciplinary
Socio Economic Objective 970106 Expanding Knowledge in the Biological Sciences
HERDC Research category C1 Refereed article in a scholarly journal
Copyright notice ©2016, American Chemical Society
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