Glycosylation is an important type of chemical modification of proteins and lipids and plays essential roles in regulating their structures, properties and functions. Cell-surface glycans are closely related to the intercellular and cell-matrix interactions and mediate cell differentiation, proliferation and migration. The progression of a variety of diseases exhibits well-established relationships with changes in the structures and expression levels of cell-surface glycans. In this context, the in-depth study of cell-surface glycans has obvious implications for understanding the intricate biological roles of cell-surface glycans in pathological processes, developing new methods for clinical diagnosis and drug screening and further revealing the essence of life. Owing to the non-template synthesis processes, complex structures and sophisticated modifications, the in situ analysis of cell-surface glycans is challenging compared with protein and nucleic acid biosensing. This chapter outlines representative analytical techniques and research advances in cell-surface glycan analysis, including cell-surface glycan recognition and labelling techniques, global analysis, competition detection format and protein-specific imaging. Through dynamic tracking of cell-surface glycans, exploration of the correlation between glycan patterns and glycan-related biological processes can be achieved.