Abstract: Objective To discuss the effect of the improved methods of reused ophthalmic microsurgery instruments and to improve the cleaning quality of the instrument for preventing infection in hospital. Methods The methodology used in this research is to distribute 600 instruments which are chosen from operation room from September to October in 2016 into two groups randomly, namely, the regular group and improved group, with 300 devices in each group. The routine cleaning method used by the regular group from sterilization and supply center (multi-enzyme ultrasonic cleaning) will be used to compare the method adopted by the improved group which comprises pretreatment (put multi-enzyme solution) ultrasound, acid cleaning agent as well as lubricant oil for medical devices cleaning. The qualified rate after cleaning, comparison of bacteriological examination and flexibility and loss rate of device will be compared between two methods. Results Although there is no significant qualified rate difference between regular group and improved group in terms of visual method (P>0.05), the qualified rate of microscopic examination in the improved group is significantly superior to the regular group (P<0.05). In addition, the improved group is better than the regular group in regard to the flexibility (P<0.05), and the loss rate of improved group is lower than that of the regular group (P<0.05). It should be noted that there is also an obvious difference between two groups in regard to positive rate of bacterial test after cleaning (P<0.05). Conclusion In the improved group, the flexibility of ophthalmic microsurgical instruments was increased by 9%, the loss rate was decreased by 2%, the bacterial test negative rate was increased by 6%, and the pass rate of microscopy was increased by 18.3%. Improved group which used pretreatment, ultrasonic and enzyme washing, acid detergent and medical lubricant can effectively improve the reuse of the ophthalmic microsurgery instrument, promote small rust stains and spots to be removed, enhance the cleaning quality the reuse of the ophthalmic microsurgery device and increase the flexibility of utilization of devices and decrease the loss rate.