This investigation used field emission in-lens scanning electron microscopy (FEISEM) for the study of tooth surfaces, with particular reference to adhesive bonding and acid conditioning. Dentin wafers with an intact enamel periphery were treated by either ethylenediaminetetraacetic acid (EDTA) (pH 7.4) or phosphoric acid (pH 0.7). The samples were then fixed, sequentially dehydrated in alcohol, and either air- or critical point-dried. After coating, surfaces were examined by FEISEM. For enamel, intraprismatic crystals were clearly recognizable, with the crystals showing both a longitudinal and parallel orientation to the long axis of the prisms. For dentin, the surface ultrastructure (mineral crystals and collagen banding) for the both untreated and treated samples was observed. Fine structures measuring on the order of 6 nm were also observed on samples treated by EDTA. We conclude that FEISEM can routinely provide high-resolution images of enamel and dentin, and that it has the capability of revealing the defined distribution of crystals and collagen fibers in dental tissues.