Visualizing Cell Type-Specific Glycan Profiles in Tissue Sections With Glycan Painting

Glycans play critical roles in numerous biological processes. Therefore, visualizing the spatial distribution of glycans within cells and tissues is essential for understanding tissue architecture and cellular identity. Glycan painting is a novel tissue-staining method that employs three fluorescently labeled lectins with different glycan-binding specificities to visualize cell type-specific glycan profiles as unique color patterns. In this study, the protocol was validated using formalin-fixed paraffin-embedded sections of mouse embryos. Although Glycan painting is a qualitative method, each cell population that is difficult to distinguish with a single lectin can be differentiated as unique color patterns generated from the combined images of three fluorescent lectins. Moreover, because it does not require costly reagents or specialized equipment, Glycan painting provides a cost-effective and broadly accessible approach for full-color tissue imaging and cell-type discrimination. Key features • Glycan painting enables full-color visualization of cell type-specific glycan profiles through simultaneous staining with three fluorescently labeled lectins with different glycan-binding abilities. • Applicable to formalin-fixed paraffin-embedded tissue sections. • Provides comprehensive visualization of tissue architecture while simultaneously facilitating cell-type discrimination. • Compatible with standard fluorescence microscopy and uses readily available lectins and reagents, enabling rapid adoption without specialized instrumentation.