Detailed knowledge of dentine structure, and especially that of the dentinal tubules, is essential in order to understand dentine permeability and to interpret data from investigations on dentine adhesive materials. The aim here was to examine the density and branching of dentinal tubules in human teeth by light and scanning electron microscopy. Stained and unstained demineralized sections and undemineralized fractured specimens were studied. Statistically significant differences in the density of tubules were found depending on location. Differences in density of tubules between the peripheral and inner aspects were more marked in the crown than in root. The mean number of dentinal tubules in the middle part of the root was significantly lower than in the middle part of the crown. The density of the tubules in the outer dentine at the cusp location was also significantly different from that subjacent to the occlusal fissure. The number of branches of dentinal tubules was particularly abundant in locations where the density of tubules was low. The branching patterns revealed an intricate and profuse canalicular, anastomosing system, criss-crossing the intertubular dentine. Three types of branches, major, fine and microbranches, were identified on the basis of size, direction and location. Major branches, 0.5-1.0 micron dia., were the typical delta branchings found peripherally. Fine branches, 300-700 nm dia., forked off at 45 degrees and were abundant in areas such as in the root where the density of the tubules was relatively low. Microbranches, 25-200 nm dia., extended at right angles from the tubules in all parts of the dentine. The findings emphasize the need for detailed characterization of dentine substrates for adhesive testing and of samples used in permeability studies.