Control of film structure in multi-step stretching process is complicated because orientation and relaxation of polymer chains during the preceding steps of stretching history affects the structure formation behavior in the subsequent stretching processes. In this research, stress-optical behavior of PS films during two-step uni-axial stretching and relaxation processes with various intermittent holding period was investigated near the glass transition temperature. Simultaneous on-line measurements of stress and birefringence were performed to clarify the characteristics of the oriented structure of various relaxation times developed in the amorphous structure. Particular attention was paid to the effect of the period of relaxation after a certain level of stretching on the structure formation behavior in the subsequent stretching process. At a relatively high temperature, stress versus birefringence behavior simply followed the stress-optical rule during stretching, holding, re-stretching and holding processes. The stress and birefringence levels increased with the decrease of temperature. When the temperature was further lowered and became close enough to the glass transition temperature, sudden increase and reduction of stress were observed immediately after the starting and stopping of the stretching, respectively. In this case, hysteresis was observed in the stress versus birefringence relation, in which stress-optical rule was hold during the relaxation process. When the intermittent holding period was changed, the stress versus birefringence relation basically followed the same hysteresis behavior except for the region of transition between stretching and relaxation. It is interesting to note that the tensile modulus at the initial stretching process was about ten times lower than that observed at the re-stretching process, whereas the modulus at the re-stretching process was virtually constant with the change in the holding period.