Abstract We investigated whether workers of the ant species Lasius niger are able to sense and discriminate the coarseness of the substrate on which they walk. First, we studied the way in which substrate coarseness affects the ants’ locomotory behaviour. Second, we investigated the spontaneous preference of ants for substrates of different coarseness. And third, we tested with a differential conditioning procedure the ants’ capacity to learn to associate a given coarseness with a food reward. The locomotory behaviour of ants differed according to substrate coarseness: ants moved significantly faster and had more sinuous trajectories on a fine than on a coarse substrate. No spontaneous preference for a substrate of a given coarseness was observed and, even after 20 successive conditioning trials, there was little evidence of the effect of experience on substrate coarseness discrimination. Overall however, ants trained on fine sand made significantly more correct choice than those trained on coarse sand. We discuss these results and argue that in L. niger substrate coarseness may be more important at the collective level, by interacting with the chemical properties of the pheromone trail used in mass recruitment to food source, than at the individual level.