A bioluminescent Pseudomonas aeruginosa was incorporated into an in vitro static diffusion method to determine whether light output could be used as a measure of wound dressing efficacy. A significant linear correlation was observed between viable counts and bioluminescence during exponential growth in planktonic culture (r(2) = 0.969). Exponential-phase cells were used to inoculate cellulose discs for integration into an in vitro wound model that incorporated a reservoir of serum. A significant linear correlation was found between bioluminescence (photon counts monitored by a low-light camera) and viable counts in this growth environment (r(2) = 0.982). Three antimicrobial wound dressings were applied to the surface of freshly prepared sample discs within the wound model, and the kill kinetics were codetermined by photon and viable counts. Quantifiable kill rates gave the same order of efficacy for the three wound dressings using both types of measurement, and a significant linear correlation was shown between photon and viable counts (r(2) = 0.873) within this killing environment. Under all defined conditions, a significant linear correlation between bioluminescence and viable counts was shown but the actual slope of the correlation was different, depending on the physicochemical environment of the cells. Hence, significantly more light per cell (P < 0.0001) was produced when cells in discs were exposed to a killing environment compared to a growing environment. As long as defined conditions are employed, the resulting linear correlation enables the state of the system to be continually monitored without disturbance, allowing more immediate and accurate calculations of kill rates without the need for viable counting.