Visualizing Cell Type–Specific Glycan Profiles in Tissue Sections With Glycan Painting
- 1Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, Japan
Abstract
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.
Related
- Glycan reachability analysis: A bottleneck-aware framework for inferring tissue-specific glycan biosynthetic potential from transcriptomics
- High-content morphological profiling by Cell Painting in 3D spheroids
- Glycan-mediated binding to collagen by a C-type lectin-like domain of the endocytic receptor Endo180 / uPARAP
- Cell Painting Identifies Phenotypic Clusters Associated with Anticancer Drug Responses in OVCAR-3 Ovarian Cancer Cells
- Loss of the Asn333 high-mannose N-glycan impairs ACP3 folding and drives disease-associated dysfunction
BibTeX
@article{nagasaki2026visualizing,
title = {{Visualizing Cell Type–Specific Glycan Profiles in Tissue Sections With Glycan Painting}},
author = {Akira Nagasaki and Yoichi Shinkai},
journal = {Bio-protocol},
year = {2026},
month = oct,
doi = {10.21769/bioprotoc.5859},
url = {https://doi.org/10.21769/bioprotoc.5859},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Visualizing Cell Type–Specific Glycan Profiles in Tissue Sections With Glycan Painting
AU - Akira Nagasaki
AU - Yoichi Shinkai
PY - 2026
DA - 2026/10/20
JO - Bio-protocol
DO - 10.21769/bioprotoc.5859
UR - https://doi.org/10.21769/bioprotoc.5859
AB - 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.
SN - 2331-8325
ER -