Glycosylation, a fundamental biological process, involves the attachment of glycans to proteins, lipids, and RNA, and plays a crucial role in various biological pathways. It is of great significance to obtain the precise spatial distribution of glycosylation modifications at cellular and tissue levels. Herein, we introduce LectoScape, a novel method enabling detailed imaging of the glycome in tissues at a resolution of up to 1 m through imaging mass cytometry (IMC). The selection of 12 non-overlapping lectins based on microarray technology allows for the multiplexed detection of various glycans. Moreover, we have developed an efficient labeling strategy for these lectins. Significantly, our approach enables the simultaneous imaging of a diverse range of glycan motifs from N-glycan, O-glycan, and glycolipid, which is not achievable by other technology. Through the application of LectoScape, we have successfully identitied distinct glycan structures in various cell types, leading to a deeper understanding of glycan distribution across human tissues. Remarkably, our method has identified α2,3 sialic acid and terminal GalNAc as diagnostic indicators for cervical intrepithelial neoplasia, showcasing the potential of this technique in cancer diagnosis by detecting abnormal glycosylation.