Due to the wide presence of carbohydrates in nature and their crucial roles in numerous important biological processes, oligosaccharides have attracted a lot of attention in synthetic organic chemistry community. Many innovative synthetic methods have been developed for oligosaccharide synthesis, among which the pre‐activation‐based glycosylation is particularly noteworthy. Traditionally, glycosylation reactions are carried out when the glycosyl donor and the acceptor are both present when the promoter is added. In comparison, the pre‐activation‐based glycosylation is unique, where the glycosyl donor is activated by the promoter in the absence of the acceptor. Upon complete donor activation, the acceptor is added to the reaction mixture enabling glycosylation. The key step in any oligosaccharide synthesis is the stereoselective formation of the glycosidic bond. As donor activation and acceptor glycosylation are temporally separated, pre‐activation‐based glycosylation can bestow unique stereochemical control. This review systematically discusses factors impacting the stereochemical outcome of a pre‐activation‐based glycosylation reaction including substituents on the glycosyl donor, reaction solvent, and additives. Applications of pre‐activation‐based stereoselective glycosylation in assembly of complex oligosaccharides are also discussed.