Artificial microcosm plaques were grown in a five-plaque culture system for up to 6 weeks, reaching a maximum depth of several mm. Procedures for long-term pH measurement with glass electrodes were established; they showed that the application of 5 or 10% sucrose for 6 min with a slow continuous flow of a basal medium containing mucin (BMM) generated the pH changes characteristic of in vivo Stephan curves. These pH responses were reproducible between plaques. Plaque mass and thickness were critical variables. Successive, sucrose-induced pH curves in plaques up to 4 mm thickness showed minor reductions only in the amplitude and rates of pH change. In plaques over 4 mm thick there was a pronounced reduction in pH response to successive sucrose applications, indicating increased diffusion limitations--a result of plaque growth to seal in the freshly-inserted pH electrode. In plaques of 6 mm maximum thickness, 10% sucrose induced a decrease to below pH 5.5 lasting 24 h, compared to the pH response in 2 mm thick plaque, which returned to the resting pH in 2 h. Differences in pH of up to 0.9 units were identified in thick plaques between inner and outer layers. The BMM flow rate was a critical determinant of the amplitude of the pH response to sucrose and subsequent return to resting pH. These results confirm, for microcosm plaque, the importance of clearance dynamics and diffusion-limited gradients in regulating plaque pH.