In this research, we developed a variable volume balloon sensor for minimally invasive surgery. But in the process of manufacturing the balloon sensors, generation of individual difference of each balloon characteristic is inevitable. To solve this problem, in this paper, we propose a initialization method of balloon sensors. The method is consisted of two processes. One is balloon material stabilization. Molecules of Silicone are fatigued by cyclic stress, so several times balloon construction by the auto pressure controller is effective. Another is balloon sensor value calibration. Adequate zero point of balloon sensor value is able to be estimated by balloon characteristic. We did an experiment of the method of balloon initialization, and result shows effectiveness of balloon stabilization and calibration.