The lag time of single cells of Listeria innocua grown on the surface of Brain Heart Infusion Agar was studied by microscopy and image analysis. An experimental set-up that enabled relocation of the cells on the agar surface was developed and used to collect data from 50 to 100 individual cells at a time. Reuterin was added at different concentrations (0-10[no-break space]AU/ml) and it was observed that it increased both the lag time of the cells and its variance. Furthermore, for a large proportion of cells, reuterin completely prevented the cell division within the time of observation. Reuterin in combination with low pH inhibited the cell division even more efficiently. A similar effect was observed for the combination of reuterin and sodium chloride. Our experimental set-up provides a good model system for generating data on the lag time of single cells on solid surfaces, which can improve the predictions of microbial growth on solid food matrices.