This chapter describes recent advances in the preparations, characterization, and evaluation of glycoliposomes and their biomedical research and applications. Many strategies for surface glyco-functionalization of liposomes have been reported in the past decades, of which two strategies/approaches have been commonly used. The first approach, named as direct liposome formulation approach, is to synthesize the glycolipid ligand first, followed by the preparation of liposomes in combination with other principal lipids. The second approach, named as postfunctionalization approach, mainly involves grafting the carbohydrate ligand onto the preformed liposomes via various bioconjugation chemistries. Cell surface carbohydrates have specific interactions with proteins, which play an important role in various biological recognition processes such as fertilization, metastasis, inflammations, and host–pathogen adhesion, thereby serving as attractive molecules for surface modification of liposomes with purpose for specific biomedical applications. Recent advances in the preparations, characterization, and evaluation of glyco-functionalized liposomes speed up glycoliposome research and applications.