It is hoped that the level of microgravity can be evaluated in scientific research. At present, part of the research work of microgravity measurement is carried out by laser interference method. Because the characteristics of laser instrument will be seriously limited by manual adjustment, it will lead to more time-consuming and uneven initial error before each measurement. Aiming at this phenomenon, this paper proposes a single frequency laser spot center recognition and analysis algorithm. By using the characteristics of Gaussian spot image connected region the automatic alignment algorithm of single frequency laser spot interference is realized, which can improve the alignment of interference spot, reduce the measurement error, and improve the precision of microgravity measurement. Finally, the algorithm is verified in the simulation environment and the actual measurement environment, and the results show that the measurement error of microgravity acceleration can be reduced by 2.199*10-7 under different strategies in the simulation environment, and the accuracy can be improved by about 12.8%; The average time of automatic adjustment is about 10s, which is much faster than the actual adjustment time. Finally, the error in the laser interferometric acceleration measurement optical path instrument can be controlled within 50μm to complete the automatic interference closed-loop control adjustment of the optical spot, which verifies the feasibility of the algorithm.