MOTIVATION: Cell membrane segmentation in electron microscopy (EM) images is a crucial step in EM image processing. However, while popular approaches have achieved performance comparable to that of humans on low-resolution EM datasets, they have shown limited success when applied to high-resolution EM datasets. The human visual system, on the other hand, displays consistently excellent performance on both low and high resolutions. To better understand this limitation, we conducted eye movement and perceptual consistency experiments. Our data showed that human observers are more sensitive to the structure of the membrane while tolerating misalignment, contrary to commonly used evaluation criteria. Additionally, our results indicated that the human visual system processes images in both global-local and coarse-to-fine manners. RESULTS: Based on these observations, we propose a computational framework for membrane segmentation that incorporates these characteristics of human perception. This framework includes a novel evaluation metric, the perceptual Hausdorff distance (PHD), and an end-to-end network called the PHD-guided segmentation network (PS-Net) that is trained using adaptively tuned PHD loss functions and a multiscale architecture. Our subjective experiments showed that the PHD metric is more consistent with human perception than other criteria, and our proposed PS-Net outperformed state-of-the-art methods on both low- and high-resolution EM image datasets as well as other natural image datasets. AVAILABILITY AND IMPLEMENTATION: The code and dataset can be found at https://github.com/EmmaSRH/PS-Net.