Objective: To provide the basis for the clinical development of ultrasonic shoulder preparation by comparing the roughness and three-dimensional topography of shoulder surface between ultrasonic instruments and conventional rotary instruments, to make preliminary suggestions for clinical use of ultrasonic instruments.Methods: (1) Four areas of buccal and palatal surfaces of six extracted human premolars were prepared with different grit size of rotary instruments. Polyether was used to take impression of the shoulder area, 3-D topography measurement laser microscope (3-D TMLM) was used to scan the impressions and compare the shoulder surface roughness of the four areas. (2) Six extracted human premolars were prepared, mesial half of the shoulder was finished with traditional rotary instruments and distal half with ultrasonic instruments. Polyether was used to take impression of the shoulder area, 3-D TMLM was used to scan the impressions and compare the shoulder surface roughness and 3-D topography, and the shoulder surface morphology was observed by surgical microscope (×25 magnification). (3) Twenty extracted human maxillary symmetrical homonymous anterior teeth were poured into die stone using artificial gingiva, ultrasonic instruments group and rotary instruments group were divided randomly. After preparing the teeth and taking the shoulder impression with polyether in dental simulate on the training system, the surface roughness of the shoulder impression in mesial, middle and distal areas was scanned and compared. The data were analyzed using SPSS 25.0 software package.Results: (1) There was no significant difference in Ra and Rz values between the abutment shoulder and impression shoulder in different areas. (2) The surface roughness of the shoulder impression prepared by ultrasonic instruments [Ra:(6.59±2.33) μm, Rz:(34.69±7.29) μm] was significantly smaller than that of the rotary instruments [Ra:(21.79±4.89) μm, Rz:(91.69±14.82) μm] (P<0.05). The morphology of the shoulder prepared by ultrasonic instruments was clear and continuous under microscope observation. (3) The surface roughness of each area of the shoulder prepared by ultrasonic instruments was significantly lower than that of the rotary instruments (P<0.001); there was no significant difference of the surface roughness (Ra) in each area of the shoulder impression after ultrasonic instrument preparation, while the shoulder impression roughness in the mesial and distal areas was significantly higher than that in the middle area (P<0.001) after rotary instrument preparation.Conclusion: Compared with the rotary instruments, the ultrasonic instruments can obtain a smoother shoulder surface, especially can significantly improve the shoulder preparation effect near the proximal surface.