FIG. 5.

Effect of lytSR on biofilm maturation. S. aureus UAMS-1 (parent), KB999 (lytS mutant), and KB999(pBK5) (complemented strain) biofilms were grown in flow-cell chambers and analyzed after 3 days of growth. (A) Representative biofilm macro images were taken using a Canon EOS 350D digital camera. (B) The samples were stained with Syto-9 (green) and Toto-3 (red) to indicate live and dead cell populations, respectively, and visualized by CLSM equipped with a Plan-Apochromat 10× objective. The images presented are top-down views of each biofilm and are accompanied by orthogonal views (above). (C) Representative z-stack images from each biofilm were compiled, and the biofilm adjacent to the substratum is visualized as a bottom-up view. (D) The biofilm images were analyzed using COMSTAT software, and total biomass (black bars; units = μm³/μm²), average thickness (white bars; units = μm), and average colony volume at the substratum (gray bars; unit = 10 nm³) for each strain are presented. The error bars correspond to the standard errors of the means.