Evaluation of environmental scanning electron microscopy for analysis of Proteus mirabilis crystalline biofilms in situ on urinary catheters

Nina Holling, Cinzia Dedi, Caroline E Jones, Joseph A Hawthorne, Geoffrey W Hanlon, Jonathan P Salvage, Bhavik A Patel, Lara M Barnes, Brian V Jones

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Proteus mirabilis is a common cause of catheter-associated urinary tract infections and frequently leads to blockage of catheters due to crystalline biofilm formation. Scanning electron microscopy (SEM) has proven to be a valuable tool in the study of these unusual biofilms, but entails laborious sample preparation that can introduce artefacts, undermining the investigation of biofilm development. In contrast, environmental scanning electron microscopy (ESEM) permits imaging of unprocessed, fully hydrated samples, which may provide much insight into the development of P. mirabilis biofilms. Here, we evaluate the utility of ESEM for the study of P. mirabilis crystalline biofilms in situ, on urinary catheters. In doing so, we compare this to commonly used conventional SEM approaches for sample preparation and imaging. Overall, ESEM provided excellent resolution of biofilms formed on urinary catheters and revealed structures not observed in standard SEM imaging or previously described in other studies of these biofilms. In addition, we show that energy-dispersive X-ray spectroscopy (EDS) may be employed in conjunction with ESEM to provide information regarding the elemental composition of crystalline structures and demonstrate the potential for ESEM in combination with EDS to constitute a useful tool in exploring the mechanisms underpinning crystalline biofilm formation.

Original languageEnglish
Pages (from-to)20-7
Number of pages8
JournalFEMS Microbiology Letters
Volume355
Issue number1
DOIs
Publication statusPublished - 1 Jun 2014

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Proteus mirabilis
Urinary Catheters
Biofilms
Electron Scanning Microscopy
X-Ray Emission Spectrometry
Catheter-Related Infections
Urinary Tract Infections
Artifacts
Catheters

Keywords

  • Biofilms/growth & development
  • Microscopy, Electron, Scanning/methods
  • Proteus mirabilis/physiology
  • Urinary Catheters/microbiology

Cite this

Evaluation of environmental scanning electron microscopy for analysis of Proteus mirabilis crystalline biofilms in situ on urinary catheters. / Holling, Nina; Dedi, Cinzia; Jones, Caroline E; Hawthorne, Joseph A; Hanlon, Geoffrey W; Salvage, Jonathan P; Patel, Bhavik A; Barnes, Lara M; Jones, Brian V.

In: FEMS Microbiology Letters, Vol. 355, No. 1, 01.06.2014, p. 20-7.

Research output: Contribution to journalArticle

Holling, Nina ; Dedi, Cinzia ; Jones, Caroline E ; Hawthorne, Joseph A ; Hanlon, Geoffrey W ; Salvage, Jonathan P ; Patel, Bhavik A ; Barnes, Lara M ; Jones, Brian V. / Evaluation of environmental scanning electron microscopy for analysis of Proteus mirabilis crystalline biofilms in situ on urinary catheters. In: FEMS Microbiology Letters. 2014 ; Vol. 355, No. 1. pp. 20-7.
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AU - Salvage, Jonathan P

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AB - Proteus mirabilis is a common cause of catheter-associated urinary tract infections and frequently leads to blockage of catheters due to crystalline biofilm formation. Scanning electron microscopy (SEM) has proven to be a valuable tool in the study of these unusual biofilms, but entails laborious sample preparation that can introduce artefacts, undermining the investigation of biofilm development. In contrast, environmental scanning electron microscopy (ESEM) permits imaging of unprocessed, fully hydrated samples, which may provide much insight into the development of P. mirabilis biofilms. Here, we evaluate the utility of ESEM for the study of P. mirabilis crystalline biofilms in situ, on urinary catheters. In doing so, we compare this to commonly used conventional SEM approaches for sample preparation and imaging. Overall, ESEM provided excellent resolution of biofilms formed on urinary catheters and revealed structures not observed in standard SEM imaging or previously described in other studies of these biofilms. In addition, we show that energy-dispersive X-ray spectroscopy (EDS) may be employed in conjunction with ESEM to provide information regarding the elemental composition of crystalline structures and demonstrate the potential for ESEM in combination with EDS to constitute a useful tool in exploring the mechanisms underpinning crystalline biofilm formation.

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