Applying the Mesolens to microbiology

investigating the structural organisation of bacterial biofilms

Research output: Contribution to conferencePoster

65 Downloads (Pure)

Abstract

Biofilms pose a public health risk due to their ability to protect bacteria from mechanical, environmental and chemical factors. Thereby they can confer resistance to their constituent bacteria and serve as a vehicle for spread of antimicrobial resistance [1]. Understanding the structure of bacterial communities is critical to developing novel methods of biofilm eradication. Current techniques for imaging live biofilms are limited by sacrificing the size of the imaging volume or spatial resolution. Common approaches to imaging biofilm architecture include electron microscopy techniques [2], single or multi-photon confocal microscopy [3] or widefield epi-fluorescence microscopy using low-magnification, low-numerical aperture lenses [4]. Here we use the Mesolens, an optical microscope with a unique combination of a low magnification (x4) and a high numerical aperture (0.47) which can image specimens up to 6x6x3 mm in volume with a lateral resolution of 700 nm and an axial resolution of 7 μm [5]. Using the Mesolens, it is possible to image whole live colony biofilms with cellular resolution in a single dataset. We report the finding of intra-colony channels (measuring ca.15 μm in diameter) which form when Escherichia coli colonies are grown on a solid surface as an inherent property of biofilm formation. By tracking the movement of 200 nm fluorescent microspheres, we observe translocation of the microspheres from the basal surface of the biofilm into the colony with specific localisation to the channel systems. The uptake of microspheres by the colony infers that these features are inherent to biofilm formation and provide a role in structural support. The biofilms in this work were grown on a nutrient-rich solid medium, and by expanding from the observations of our bead uptake assay we can deduce that the channels may also play a role in nutrient uptake and dissemination throughout the colony. These findings serve as evidence of a fundamental principle of structural biology and bacterial organisation.
Original languageEnglish
Publication statusPublished - 27 Jun 2018
EventRMS Frontiers in Bioimaging 2018 - TIC, University of Strathclyde, Glasgow, United Kingdom
Duration: 27 Jun 201828 Jun 2018
https://www.rms.org.uk/discover-engage/event-calendar/frontiers-in-bioimaging-2018.html

Conference

ConferenceRMS Frontiers in Bioimaging 2018
CountryUnited Kingdom
CityGlasgow
Period27/06/1828/06/18
Internet address

Fingerprint

Biofilms
Microbiology
Microspheres
Bacteria
Bacterial Structures
Photons
Fluorescence Microscopy
Confocal Microscopy
Lenses
Electron Microscopy
Public Health
Escherichia coli
Food

Keywords

  • Mesolens
  • large field of view
  • optical microscopy
  • biofilm
  • E. coli
  • biomedical Imaging
  • spatial patterning

Cite this

Rooney, L. M., McCann, L., Hoskisson, P. A., & McConnell, G. (2018). Applying the Mesolens to microbiology: investigating the structural organisation of bacterial biofilms. Poster session presented at RMS Frontiers in Bioimaging 2018, Glasgow, United Kingdom.
Rooney, Liam M. ; McCann, Lee ; Hoskisson, Paul A. ; McConnell, Gail. / Applying the Mesolens to microbiology : investigating the structural organisation of bacterial biofilms. Poster session presented at RMS Frontiers in Bioimaging 2018, Glasgow, United Kingdom.
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author = "Rooney, {Liam M.} and Lee McCann and Hoskisson, {Paul A.} and Gail McConnell",
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note = "RMS Frontiers in Bioimaging 2018 ; Conference date: 27-06-2018 Through 28-06-2018",
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Rooney, LM, McCann, L, Hoskisson, PA & McConnell, G 2018, 'Applying the Mesolens to microbiology: investigating the structural organisation of bacterial biofilms' RMS Frontiers in Bioimaging 2018, Glasgow, United Kingdom, 27/06/18 - 28/06/18, .

Applying the Mesolens to microbiology : investigating the structural organisation of bacterial biofilms. / Rooney, Liam M.; McCann, Lee; Hoskisson, Paul A.; McConnell, Gail.

2018. Poster session presented at RMS Frontiers in Bioimaging 2018, Glasgow, United Kingdom.

Research output: Contribution to conferencePoster

TY - CONF

T1 - Applying the Mesolens to microbiology

T2 - investigating the structural organisation of bacterial biofilms

AU - Rooney, Liam M.

AU - McCann, Lee

AU - Hoskisson, Paul A.

AU - McConnell, Gail

PY - 2018/6/27

Y1 - 2018/6/27

N2 - Biofilms pose a public health risk due to their ability to protect bacteria from mechanical, environmental and chemical factors. Thereby they can confer resistance to their constituent bacteria and serve as a vehicle for spread of antimicrobial resistance [1]. Understanding the structure of bacterial communities is critical to developing novel methods of biofilm eradication. Current techniques for imaging live biofilms are limited by sacrificing the size of the imaging volume or spatial resolution. Common approaches to imaging biofilm architecture include electron microscopy techniques [2], single or multi-photon confocal microscopy [3] or widefield epi-fluorescence microscopy using low-magnification, low-numerical aperture lenses [4]. Here we use the Mesolens, an optical microscope with a unique combination of a low magnification (x4) and a high numerical aperture (0.47) which can image specimens up to 6x6x3 mm in volume with a lateral resolution of 700 nm and an axial resolution of 7 μm [5]. Using the Mesolens, it is possible to image whole live colony biofilms with cellular resolution in a single dataset. We report the finding of intra-colony channels (measuring ca.15 μm in diameter) which form when Escherichia coli colonies are grown on a solid surface as an inherent property of biofilm formation. By tracking the movement of 200 nm fluorescent microspheres, we observe translocation of the microspheres from the basal surface of the biofilm into the colony with specific localisation to the channel systems. The uptake of microspheres by the colony infers that these features are inherent to biofilm formation and provide a role in structural support. The biofilms in this work were grown on a nutrient-rich solid medium, and by expanding from the observations of our bead uptake assay we can deduce that the channels may also play a role in nutrient uptake and dissemination throughout the colony. These findings serve as evidence of a fundamental principle of structural biology and bacterial organisation.

AB - Biofilms pose a public health risk due to their ability to protect bacteria from mechanical, environmental and chemical factors. Thereby they can confer resistance to their constituent bacteria and serve as a vehicle for spread of antimicrobial resistance [1]. Understanding the structure of bacterial communities is critical to developing novel methods of biofilm eradication. Current techniques for imaging live biofilms are limited by sacrificing the size of the imaging volume or spatial resolution. Common approaches to imaging biofilm architecture include electron microscopy techniques [2], single or multi-photon confocal microscopy [3] or widefield epi-fluorescence microscopy using low-magnification, low-numerical aperture lenses [4]. Here we use the Mesolens, an optical microscope with a unique combination of a low magnification (x4) and a high numerical aperture (0.47) which can image specimens up to 6x6x3 mm in volume with a lateral resolution of 700 nm and an axial resolution of 7 μm [5]. Using the Mesolens, it is possible to image whole live colony biofilms with cellular resolution in a single dataset. We report the finding of intra-colony channels (measuring ca.15 μm in diameter) which form when Escherichia coli colonies are grown on a solid surface as an inherent property of biofilm formation. By tracking the movement of 200 nm fluorescent microspheres, we observe translocation of the microspheres from the basal surface of the biofilm into the colony with specific localisation to the channel systems. The uptake of microspheres by the colony infers that these features are inherent to biofilm formation and provide a role in structural support. The biofilms in this work were grown on a nutrient-rich solid medium, and by expanding from the observations of our bead uptake assay we can deduce that the channels may also play a role in nutrient uptake and dissemination throughout the colony. These findings serve as evidence of a fundamental principle of structural biology and bacterial organisation.

KW - Mesolens

KW - large field of view

KW - optical microscopy

KW - biofilm

KW - E. coli

KW - biomedical Imaging

KW - spatial patterning

UR - https://www.rms.org.uk/discover-engage/event-calendar/frontiers-in-bioimaging-2018.html

M3 - Poster

ER -

Rooney LM, McCann L, Hoskisson PA, McConnell G. Applying the Mesolens to microbiology: investigating the structural organisation of bacterial biofilms. 2018. Poster session presented at RMS Frontiers in Bioimaging 2018, Glasgow, United Kingdom.