This paper proposes a method combining segmentation and Gaussian fitting based on the analysis of image features to extract the center of marker under the perspective of infrared camera. Firstly, the ROI rough positioning of the marker is carried out based on the gray difference of the marker in the image, and the influence of background reflection on the segmentation is eliminated. Then, the K-means clustering algorithm is used to segment the target image. Finally, the improved Gaussian fitting extraction method is used to obtain the center coordinate of the marker spot by removing the central redundant information. The experimental results show that compared with the centroid method, Hough transform method, and circular fitting method, the algorithm in this paper has a stability error of less than 0.1 pixels for center extraction, with high repeatability and stable accuracy. Even with different degrees of occlusion, it still maintains high accuracy, which is suitable for the center extraction of marker in the positioning system under the perspective of infrared camera.