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. 2022 Oct 3;119(41):e2209699119. doi: 10.1073/pnas.2209699119

Fig. 3.

Fig. 3.

Biofilm growth dynamics of interkingdom assemblages on tooth-mimetic surface. (A) Schematic diagram of the flow-cell microfluidic culture system coupled with time-lapse confocal microscopy for visualizing biofilm growth. (B) Confocal images of the initial colonizers on the surface at 0 h, and the biofilm structure after 10 h, for interkingdom assemblage (Assembl), for aggregated S. mutans (Agg S.m.), and for fungicide-treated assemblage (250 µg/mL nystatin for 30 min). Green, S. mutans; cyan, C. albicans. (Scale bar, 100 μm.) (C) Time-resolved biofilm biovolume of S. mutans during the biofilm development. (Inset) Time-resolved biovolume of each S. mutans and C. albicans within the assemblage. Lines correspond to mean, shaded region to SD of n = 4 independent replicates. (D) Quantification of the dynamics of biofilm surface spreading. Lines correspond to mean, shaded region to SD of n = 4 independent replicates. *P < 0.05 by one-way analysis of variance with Tukey’s multiple-comparison test (t = 6.5 h). (E) Confocal image time series showing merging behavior (yellow arrowhead) of multiple individually developing interkingdom assemblages on the tooth-mimetic surface. Green, S. mutans; cyan, C. albicans. (Scale bar, 100 μm.)