Temporospatial imaging of bacterial biofilms.
Bacterial biofilms play a role in human disease by providing a protected environment for pathogens by changing the growth characteristics of the bacteria and by excluding antibiotics and host immune factors. Biofilms that form in non-sterile body sites are typically polymicrobial. These polymicrobial biofilms, such as those found in dental plaque or growing on the surface of the nasopharyngeal mucosa are shaped by a complex array of inter-species interactions. The distribution of strains and species within the biofilm structure is dictated by the offsetting influences of antagonistic and mutualistic interactions. Most of our existing knowledge of how these structures form is derived from end-point imaging or sampling of biofilms. These approaches provide limited information on how intra and inter species interactions shape the biofilm structure over time. In this work, we propose to use a novel method of time lapse imaging to visualize the development of mono- and polymicrobial biofilms in three dimensions over time. Furthermore, we envisage that we will be able to visualize the entire biofilm architecture as well as the spatial position of a specific species simultaneously over time. We will examine the influence of coaggregation, bacteriocin production and quorum sensing pathways on the architecture and species distribution of biofilms over time using our established models of polymicrobial dental and nasopharyngeal biofilms.