High-resolution novel method for tracking bacteria in a multi-species biofilm
The aim of this study is to establish a novel high resolution tracking ability of a specific bacterium in multispecies biofilm. A periodontal multispecies biofilm was constructed with Streptococcus sanguis, Actinomyces naeslundii, Porphyromonas gingivalis and Fusobacterium nucleatum. A single species was stained with fluorescein isothiocyanate (FITC). The mature biofilm was stained for viability (propidium iodide) and analysis was performed with flow cytometry. The sensitivity of the assay was compared with colony forming units (CFU) counts. A single cell suspension of P. gingivalis was grown in broth and biofilm to identify the location of these events on side scatter and forward scatter. The sensitivity of the assay was comparable to that of the CFU counts. The assay allows quantification of the ratio of a single bacterium within the biofilm, and its viable proportion. The described method is reproducible and of high resolution, and allows the examination of microbes’ composition and viability within a biofilm structure.
KeywordsMultispecies biofilm Flow cytometry Bacterial viability Periodontitis
The study was self-supported.
Compliance with ethical standards
Conflict of interest
We hereby declare that this is an original manuscript, has not been sent elsewhere, do not have commercial or other associations that might pose a conflict of interest. All authors have contributed to, seen, and approved the final submitted version of the manuscript. The study was self-supported.
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