Abstract: In order to study the method of improving the success rate of patch clamp experiment, taking the whole cell patch clamp electrophysiological detection experiment of pc12 cells as an example, by analyzing the influencing factors and controlling the key parameters of the experiment, the success rate and quality of high blocking sealing and cell membrane breakage were recorded, and the optimal parameters of the experiment were finally determined through the analysis of experimental data. The experimental results show that under the experimental equipment and environment, the optimal parameters to increase the success rate are: the tip diameter of glass microelectrode is 3 μm; the plating current of Ag/AgCl electrode is 0.1 mA and the plating time is 2 hours; the negative pressure of high resistance sealing is 1~2 mL; the negative pressure of film breaking is 0.5~1 mL; and the continuation pulse of film breaking electric shock stimulation is 3 ms.