Fig. 2

Biofilm development in the microfluidic chip. Biofilm morphologies in different pore spaces (a). Biofilm cells were stained with DAPI. Schematic diagram illustrating biofilm morphology and coverage analysis (b). Biofilm thickness was calculated as the average radial distance from the biofilm perimeter to the micropillar surface, while roughness was quantified as the standard deviation of biofilm thickness on individual micropillar surfaces. Coverage represents the proportion of pore space occupied by biofilm. The biofilm thickness, roughness, and coverage in different pore environments (c). Data are presented as mean ± standard deviation. Biofilm morphology and coverage were assessed using images sampled from random areas within three independent biological replicates per pore space. Twenty-five observations per group were analyzed for biofilm thickness and roughness, while 15 observations per group were collected for biofilm coverage analysis. Different letters indicate significant differences (p < 0.05, one-way ANOVA)