In order to achieve localization adaptability of selfwalking type tomato harvesting machine fruit separation device, the fruit separation device performance was optimized with the method of response surface analysis. The oscillator spindle speed, the oscillator eccentric mass, roller rotation speed and tomato feeding volume were chosen as the influence factors, and the fruit separation rate, breakage rate and impurity rate were used as the response indexes, a multiple regression model was established based on the response surface analysis with Matlab software, and the credibility of the prediction model was analyzed by using the R2, P and the variance of the residual S2; the effects of experimental factors on test index were analyzed based on the response surface method and the influence factors and the best combination of level were achieved, and combined with the nonlinear optimization method, the optimization calculation of fruit separation rate, damage rate and impurity rate were carried out. The results showed that when the oscillator spindle speed was 342.86r/min, vibrator eccentric mass was 40kg, roller rotation speed was 22r/min and tomato feed quantity was 19kg/s, the fruit separation rate, broken rate and impurity rate were 98.33%, 3.37% and 4.17% respectively. By the experiment, tomato separation parameters got by response surface method were feasible; the research result offered the theoretical foundation and scientific basis for the further study of the processing tomato vibration separation technology, also for fruit vibration harvesting technology parameters optimization.