Glycome is defined by the glycosylation machinery with which each cell is equipped, and this differs between species. It is evident that cells show drastic change during cell progression and differentiation associated with tumorigenesis and malformation. Histochemical approaches to analyze molecular and cellular dynamics provide useful clues to answering questions about glycan functions associated with pathology. However, development of glyco-biomarker discovery will require differential glycan analysis in a number of clinical specimens where disease lesions and normal regions from the same tissue sections are compared. In this chapter, we describe a simple but powerful method using an ultrasensitive lectin microarray, which enables rapid and systematic differential glycan analysis targeting restricted regions of formalin-fixed tissue specimens, for example, one-dot sections formatted on tissue arrays. Using this advanced technology followed by an objective statistical analysis, we can select lectin probes to best fit subsequent enrichment procedures to identify target glycoproteins that discriminate diseased and normal regions in the tissue specimens.