The spacing of short-period incremental markings in dentine was measured in longitudinal ground sections and in longitudinal demineralized silver-stained sections of permanent human canines and premolars. Measurements were made (i) within 50 microm from the granular layer of Tomes (GLT), (ii) between 100 and 200 microm from the GLT, and (iii) in the axial plane of the tallest cusps. Median values for the spacing of calcospheritic lines closest to the GLT in the ground sections increased from 1.8 to 2.8 microm as the lines gradually coalesced into a laminar pattern beyond the GLT pulpally. Median values for the spacing of short-period lines in the cuspal dentine, where dentine formation is known to be fastest, were 4.1 microm. Markings in the demineralized sections were between 25 and 39% closer together, presumably due to contraction and shrinkage during specimen preparation. The spacings of short-period incremental lines measured on ground sections of non-human primate dentine (gibbon, siamang, orang) and on pig dentine, all between 100 and 200 microm from the GLT, clustered between 2.5 and 3.5 microm. Apart from gibbon dentine (in which spacings were closer together in this position than in the others), the distribution of measurements was not significantly different in pig, orang or human dentine. However, none of the data for the comparative samples presented here revealed spacings of short-period lines anywhere close to the 16 microm previously reported for circumpulpal dentine in animals. These data suggest that there may be many other animals where the mode and, to some extent, rate of dentine mineralization close to the root surface follows a common pattern. While data for the spacing of incremental markings in dentine provide no evidence for their periodicity, it is clear that the measurements made in the ground sections match the reported daily rates of mineralization at these locations, whereas those in demineralized silver-stained sections do not. Tissue shrinkage is probably a better explanation for this than the generally accepted view that they represent 12 h increments of dentine mineralization. This study provides a better basis for identifying and describing these lines, and for distinguishing them from other kinds of incremental markings in dentine.