:A detection method on the accuracy of high-speed camera sampling sequence is proposed. Pulse signals are used to control and test the luminescence of the targets,and the detected targets are recorded with high speed camera. Through analyzing images and sampling sequence,the sampling frequency and time sequence of the high speed camera are obtained and verified. A set of sampling frequency detector was designed based on this method,and the frequency and time sequence of MS50K high-speed camera were analyzed with this detector. The results show that the frequency and time sequence of MS50K high speed camera are accurate,and the proposed method can be applied to the accuracy detection of sampling frequency and time sequence. The response time of this detector reaches 1μs after modification,which can satisfy the requirements of the sampling sequence detection equipment.