The structure, distribution and formation of accessory canals in the developing and mature molar teeth of rat mandibular jaws were investigated with scanning electron microscopy and with three-dimensional image analysis using serial light-microscopic sections. In the initial stage of the accessory canal formation, most of the canals appeared in the gaps of the epithelial root sheaths formed by their approaching each other in the initial stage of the root formation. However, some of the canals appeared in the slits which may be formed by the destruction of the epithelial root sheath in the root apex regions. When the gaps and slits were invaded by blood vessels, the regions surrounding the vessels did not mineralize but became accessory canals. Usually, an accessory canal with one blood vessel connected the periodontal ligament to the dental pulp; however, in some cases, the canals were broken off midway following the destruction of the vessels.