The strap-down inertial navigation system (SINS), especially the SINS composed by dithered ring laser gyroscope (DRLG) is a kind of equipment, which providing high reliability and performance for moving vehicles. However, the mechanical dither which is used to eliminate the “Lock-In” effect can cause vibration disturbance to the INS and lead to dithering coupling problem in the inertial measurement unit (IMU) gyroscope triad, so its further application is limited. Among DRLG errors between the true gyro rotation rate and the measured rotation rate, the frequently considered one is the input axis misalignment between input reference axis which is perpendicular to the mounting surface and gyro angular rate input axis. But the misalignment angle between DRLG dither axis and gyro angular rate input axis is often ignored by researchers, which is amplified by dither coupling problem and that would lead to negative effects especially in high accuracy SINS. In order to study the problem more clearly, the concept of misalignment between DRLG dither axis and gyro angle rate input axis is researched. Considering the error of misalignment is of the order of 10-3 rad. or even smaller, the best way to measure it is using DRLG itself by means of an angle exciter as an auxiliary. In this paper, the concept of dither axis misalignment is explained explicitly firstly, based on this, the frequency of angle exciter is induced as reference parameter, when DRLG is mounted on the angle exciter in a certain angle, the projections of angle exciter rotation rate and mechanical oscillation rate on the gyro input axis are both sensed by DRLG. If the dither axis has misalignment error with the gyro input axis, there will be four major frequencies detected: the frequency of angle exciter, the dither mechanical frequency, sum and difference frequencies of the former two frequencies. Then the amplitude spectrum of DRLG output signal obtained by the using LabVIEW program. if there are only angle exciter and the dither mechanical frequencies, the misalignment may be too small to be detected, otherwise, the amplitude of the sum and difference frequencies will show the misalignment angle between the gyro angle rate input axis and the dither axis. Finally, some related parameters such as frequency and amplitude of the angle exciter and sample rate are calculated and the results are analyzed. The simulation and experiment result prove the effectiveness of the proposed method..