Abstract: To aim precisely at the target through eye tracking will become an important control method for unmanned weapons. Calibration is an important part of eye tracking, and different calibration interfaces will produce different fixation effects due to human eye observation and physiological characteristics. By changing the background of the calibration interface and such elements of the fixed point as color, shape, size, and so on, the least-squares regression method was used to establish the line of sight to estimate mapping model calibration, 5×5 evenly distributed 25 points were tested, the fixation accuracy and precision of 27 groups of calibration interfaces were compared and analyzed, and the best calibration interface was selected, to research the human eye fixation effect of calibration interface. The experimental results showed that the fixation accuracy of interface #1 was the best (0.69°±0.43°), and the fixation precision of interface #24 was the best (0.270°±0.083°). According to the criterion of fixation accuracy and precision, calibration interface No. 1 was the best calibration interface.