Multi-level thresholding image segmentation divides an image into multiple regions of interest and is a key step in image processing and image analysis. Aiming toward the problems of the low segmentation accuracy and slow convergence speed of traditional multi-level threshold image segmentation methods, in this paper, we present multi-level thresholding image segmentation based on an improved slime mould algorithm (ISMA) and symmetric cross-entropy for global optimization and image segmentation tasks. First, elite opposition-based learning (EOBL) was used to improve the quality and diversity of the initial population and accelerate the convergence speed. The adaptive probability threshold was used to adjust the selection probability of the slime mould to enhance the ability of the algorithm to jump out of the local optimum. The historical leader strategy, which selects the optimal historical information as the leader for the position update, was found to improve the convergence accuracy. Subsequently, 14 benchmark functions were used to evaluate the performance of ISMA, comparing it with other well-known algorithms in terms of the optimization accuracy, convergence speed, and significant differences. Subsequently, we tested the segmentation quality of the method proposed in this paper on eight grayscale images and compared it with other image segmentation criteria and well-known algorithms. The experimental metrics include the average fitness (mean), standard deviation (std), peak signal to noise ratio (PSNR), structure similarity index (SSIM), and feature similarity index (FSIM), which we utilized to evaluate the quality of the segmentation. The experimental results demonstrated that the improved slime mould algorithm is superior to the other compared algorithms, and multi-level thresholding image segmentation based on the improved slime mould algorithm and symmetric cross-entropy can be effectively applied to the task of multi-level threshold image segmentation.