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Imaging live bacteria at the nanoscale: comparison of immobilisation strategies

Abstract

Atomic force microscopy (AFM) provides an effective, label-free technique enabling the imaging of live bacteria under physiological conditions with nanometre precision. However, AFM is a surface scanning technique, and the accuracy of its performance requires the effective and reliable immobilisation of bacterial cells onto substrates. Here, we compare the effectiveness of various chemical approaches to facilitate the immobilisation of Escherichia coli onto glass cover slips in terms of bacterial adsorption, viability and compatibility with correlative imaging by fluorescence microscopy. We assess surface functionalisation using gelatin, poly-L-lysine, Cell-Tak™, and Vectabond®. We describe how bacterial immobilisation, viability and suitability for AFM experiments depend on bacterial strain, buffer conditions and surface functionalisation. We demonstrate the use of such immobilisation by AFM images that resolve the porin lattice on the bacterial surface; local degradation of the bacterial cell envelope by an antimicrobial peptide (Cecropin B); and the formation of membrane attack complexes on the bacterial membrane.

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Publication details

The article was received on 27 Jun 2019, accepted on 10 Oct 2019 and first published on 10 Oct 2019


Article type: Paper
DOI: 10.1039/C9AN01185D
Analyst, 2019, Accepted Manuscript
  • Open access: Creative Commons BY license
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    Imaging live bacteria at the nanoscale: comparison of immobilisation strategies

    G. Benn, A. Pyne, M. Ryadnov and B. Hoogenboom, Analyst, 2019, Accepted Manuscript , DOI: 10.1039/C9AN01185D

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