This paper sought to investigate pupil size variability, pupil size parameters in terms of time domain and frequency domain, the autonomic activity change induced by posture change, degree of sleepiness and cognitive task (math task). With a specially designed pupil image acquisition system in the dark room, these three kinds of experiments were performed to induce a dominant state of sympathetic or parasympathetic activation. Electrocardiogram and pupil size were measured in all the experiments. Based on three experiments, we calculated heart rate variability. In the pupil size analysis, we calculated the mean and standard deviation of pupil size (in time domain), and proposed several frequency bands that exhibit different autonomic activation between different sessions. The results indicate that in terms of heart rate variability, posture change exhibited significant differences but not between sleepiness level, or between cognitive task. Pupil sizes differed only during the postures. And we found some frequency bands that correlated with autonomic activation in each experiment. While heart rate variability reflects posture change that need cardiac control, pupil size variability reflects not only posture induced autonomic activation but sleepiness and cognitive load, which is processed in the brain, in time and frequency domain parameter.