Applying the mesolens to microbiology : visualising biofilm architecture and substructure

Rooney, Liam M. and McCann, Lee and Hoskisson, Paul A. and McConnell, Gail (2018) Applying the mesolens to microbiology : visualising biofilm architecture and substructure. In: 18th European Light Microscopy Initiative Meeting 2018, 2018-06-05 - 2018-06-08.

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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 wide field 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 base 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.