A new type of self-etching agent, the single-step adhesive, was developed. The purpose of this study was to investigate the difference in the ultrastructural features of enamel and dentin surfaces following application of the prototype single-step adhesive (EXM618, 3M, MN, USA). Extracted caries-free human premolars were used in this study. Occlusal enamels of teeth--to be used as cut surface specimens--were removed perpendicular to the long axis of the specimens. The mesial and distal surfaces of these teeth, on the other hand, were used as uncut surface specimens in the test. In addition, Mega Bond (Kuraray Medical Inc., Tokyo, Japan) and Xeno CFII Bond (Dentsply Sankin, Tokyo, Japan) were used as controls. After covering half of the tooth surfaces with nail varnish (for control), the other half of each surface was treated with one of the three test adhesives--EXM618, Mega Bond, or Xeno CFII Bond--according to the manufacturer's recommendations. Conditioned enamel and dentin surfaces (i.e., decalcified depth and rugged surface) were observed with a scanning confocal laser microscope (SCLM 1100, Olympus, Tokyo, Japan; henceforth abbreviated as SCLM). Based on the findings of this study, the prototype single-step adhesive EXM618 appears to be suitable for use in dental clinics.