Fig. 4: Bacterial attachment comparing different materials, inversion and flow. | Nature Communications

Fig. 4: Bacterial attachment comparing different materials, inversion and flow.

From: Combinatorial discovery of microtopographical landscapes that resist biofilm formation through quorum sensing mediated autolubrication

Fig. 4

a Representative images of P. aeruginosa wildtype stained with Syto9 fluorescent dye (green) or P. aeruginosa wild type transformed with mCherry (red; on PU only) grown for 4 h on flat, pro- or anti-attachment TopoUnits (bright field images shown in top row) moulded from PS, PU or cyclic olefin copolymer (COC). P. aeruginosa wildtype attachment on upside down oriented TopoUnits (inverted) and under flow conditions is shown for PS TopoUnits. Scale bar: 50 µm. b Quantification of normalised mean fluorescence intensity of wildtype P. aeruginosa incubated under conditions described above (PS: n = 22 TopoUnits, PS Flow: n = 4 TopoUnits, PS Inverted: n = 5 TopoUnits, COC: n = 12 TopoUnits and PU: n = 8 TopoUnits). Statistical analysis was done using a two-way ANOVA with Dunnett’s multiple comparisons test (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). P-values for 881 and 685 topographies are: PS < 1.00E−15 and <1.00E−15, PS Flow = 2.40E−06 and 6.23E−9, PS Inverted = 7.84E−06 and 2.41E−05, COC = 3.37E−13 and 7.46E−13, PU < 1.00E−15 and <1.00E−15. c Bacterial biofilm assessment: Representative images of P. aeruginosa and S. aureus biofilms on flat, pro and anti-attachment PS topographies after 24 h incubation under static conditions. Scale bar: 50 µm. d Quantification of mean normalised fluorescence intensity for wildtype P. aeruginosa and S. aureus biofilms shown in c. P. aeruginosa n = 32 TopoUnits, S. aureus n = 24 TopoUnits. Data shown in boxes extend from the 25th to 75th percentiles and lines in boxes correspond to the median values. Whiskers go down to the smallest and up to the largest values. Statistical analysis was done using a two-way ANOVA with Dunnett’s multiple comparisons test (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). P-values for 881 and 685 topographies are: P. aeruginosa = 2.22E−13 and 2.22E−13, S. aureus = 5.29E−10 and 1.08E−10. For Fig. 4a, d, the source data are provided as a Source Data file.

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