To explore the effect of pulsating vacuum cleaning sterilizer on the cleaning quality of dental handpieces.A total of 390 newly-purchased high-speed dental handpieces were covered in the study. By the random number table method, the handpieces were divided into three groups that were cleaned by different methods-Group A ( n=130), pulsating vacuum cleaning sterilizer, group B ( n=130), automatic thermal cleaning and disinfection machine, and group C ( n=130), manual cleaning. The cleaning quality, internal cavity drying and the length of cleaning time of the three groups were compared.The cleaning quality of group A (100%) was better than those of groups B (89.2%) and C (83.8%) and the length of cleaning time of group A (1.13 h) was shorter than those of group B (1.80 h) and C (2.60 h), all showing significant differences ( P0.05).The pulsating vacuum cleaning sterilizer can effectively improve the cleaning quality of dental handpieces and shorten the cleaning time. Hence, it should be extensively applied and promoted in clinic service.