Flynn, R. A. et al. Small RNAs are modified with N-glycans and displayed on the surface of living cells. Cell 184, 3109–3124.e3122 (2021).
Tarbell, J. M. & Cancel, L. M. The glycocalyx and its significance in human medicine. J. Intern. Med. 280, 97–113 (2016).
Möckl, L. The emerging role of the mammalian glycocalyx in functional membrane organization and immune system regulation. Front. Cell Dev. Biol. 8, 253 (2020).
Crocker, P. R., Paulson, J. C. & Varki, A. Siglecs and their roles in the immune system. Nat. Rev. Immunol. 7, 255–266 (2007).
Smith, B. A. H. & Bertozzi, C. R. The clinical impact of glycobiology: targeting selectins, Siglecs and mammalian glycans. Nat. Rev. Drug Discov. 20, 217–243 (2021).
Lipowsky, H. H. The endothelial glycocalyx as a barrier to leukocyte adhesion and its mediation by extracellular proteases. Ann. Biomed. Eng. 40, 840–848 (2012).
Paszek, M. J. et al. The cancer glycocalyx mechanically primes integrin-mediated growth and survival. Nature 511, 319–325 (2014).
Crouch, L. I. et al. The role of glycans in health and disease: regulators of the interaction between gut microbiota and host immune system. Semin. Immunol. 73, 101891 (2024).
Pinho, S. S., Alves, I., Gaifem, J. & Rabinovich, G. A. Immune regulatory networks coordinated by glycans and glycan-binding proteins in autoimmunity and infection. Cell. Mol. Immunol. 20, 1101–1113 (2023).
Pinho, S. S. & Reis, C. A. Glycosylation in cancer: mechanisms and clinical implications. Nat. Rev. Cancer 15, 540–555 (2015).
Bagdonaite, I. et al. Glycoproteomics. Nat. Rev. Methods Primers 2, 48 (2022).
Wu, X. et al. Imaging single glycans. Nature 582, 375–378 (2020).
Arkill, K. P. et al. 3D reconstruction of the glycocalyx structure in mammalian capillaries using electron tomography. Microcirculation 19, 343–351 (2012).
Möckl, L. et al. Quantitative super-resolution microscopy of the mammalian glycocalyx. Dev. Cell 50, 57–72.e56 (2019).
McDowell, C. T., Lu, X., Mehta, A. S., Angel, P. M. & Drake, R. R. Applications and continued evolution of glycan imaging mass spectrometry. Mass Spectrom. Rev. 42, 674–705 (2023).
Anggara, K. et al. Direct observation of glycans bonded to proteins and lipids at the single-molecule level. Science 382, 219–223 (2023).
Ebong, E. E., Macaluso, F. P., Spray, D. C. & Tarbell, J. M. Imaging the endothelial glycocalyx in vitro by rapid freezing/freeze substitution transmission electron microscopy. Arterioscler. Thromb. Vasc. Biol. 31, 1908–1915 (2011).
Chevalier, L. et al. Electron microscopy approach for the visualization of the epithelial and endothelial glycocalyx. Morphologie 101, 55–63 (2017).
Kappler, K. & Hennet, T. Emergence and significance of carbohydrate-specific antibodies. Genes Immun. 21, 224–239 (2020).
Stanley, P. Genetics of glycosylation in mammalian development and disease. Nat. Rev. Genet. 25, 715–729 (2024).
Cummings, R. D. et al. in Essentials of Glycobiology (eds Varki, A. et al.) 645–662 (Cold Spring Harbor Laboratory Press, 2022).
Laughlin, S. T., Baskin, J. M., Amacher, S. L. & Bertozzi, C. R. In vivo imaging of membrane-associated glycans in developing zebrafish. Science 320, 664–667 (2008).
Letschert, S. et al. Super-resolution imaging of plasma membrane glycans. Angew. Chem. Int. Ed. Engl. 53, 10921–10924 (2014).
Reily, C., Stewart, T. J., Renfrow, M. B. & Novak, J. Glycosylation in health and disease. Nat. Rev. Nephrol. 15, 346–366 (2019).
Aebi, M., Bernasconi, R., Clerc, S. & Molinari, M. N-Glycan structures: recognition and processing in the ER. Trends Biochem. Sci. 35, 74–82 (2010).
Rust, M. J., Bates, M. & Zhuang, X. Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Methods 3, 793–795 (2006).
Mateos-Gil, P., Letschert, S., Doose, S. & Sauer, M. Super-resolution imaging of plasma membrane proteins with click chemistry. Front. Cell Dev. Biol. 4, 98 (2016).
Riera, R. et al. Single-molecule imaging of glycan–lectin interactions on cells with Glyco-PAINT. Nat. Chem. Biol. 17, 1281–1288 (2021).
Helmerich, D. A. et al. Photoswitching fingerprint analysis bypasses the 10-nm resolution barrier. Nat. Methods 19, 986–994 (2022).
Reinhardt, S. C. M. et al. Angstrom-resolution fluorescence microscopy. Nature 617, 711–716 (2023).
Laughlin, S. T. & Bertozzi, C. R. Metabolic labeling of glycans with azido sugars and subsequent glycan-profiling and visualization via Staudinger ligation. Nat. Protoc. 2, 2930–2944 (2007).
Scinto, S. L. et al. Bioorthogonal chemistry. Nat. Rev. Methods Primers 1, 30 (2021).
Jungmann, R. et al. Single-molecule kinetics and super-resolution microscopy by fluorescence imaging of transient binding on DNA origami. Nano Lett. 10, 4756–4761 (2010).
Axelrod, D., Thompson, N. L. & Burghardt, T. P. Total internal reflection fluorescent microscopy. J. Microsc. 129, 19–28 (1983).
Heilemann, M. et al. Subdiffraction-resolution fluorescence imaging with conventional fluorescent probes. Angew. Chem. Int. Ed. Engl. 47, 6172–6176 (2008).
Sahl, S. J. et al. Direct optical measurement of intramolecular distances with angstrom precision. Science 386, 180–187 (2024).
Ester, M., Kriegel, H.-P., Sander, J. & Xu, X. A density-based algorithm for discovering clusters in large spatial databases with noise. In Proc. Second International Conference on Knowledge Discovery and Data Mining 226–231 (AAAI, 1996).
Suzuki, N., Abe, T., Hanzawa, K. & Natsuka, S. Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures. Sci. Rep. 11, 6334 (2021).
Sumer-Bayraktar, Z. et al. N-Glycans modulate the function of human corticosteroid-binding globulin. Mol. Cell. Proteomics 10, M111.009100 (2011).
Schjoldager, K. T., Narimatsu, Y., Joshi, H. J. & Clausen, H. Global view of human protein glycosylation pathways and functions. Nat. Rev. Mol. Cell Biol. 21, 729–749 (2020).
Schnaar, R. L. Glycobiology simplified: diverse roles of glycan recognition in inflammation. J. Leukoc. Biol. 99, 825–838 (2016).
Shurer, C. R. et al. Physical principles of membrane shape regulation by the glycocalyx. Cell 177, 1757–1770.e1721 (2019).
Raba, G. & Luis, A. S. Mucin utilization by gut microbiota: recent advances on characterization of key enzymes. Essays Biochem. 67, 345–353 (2023).
Chin-Hun Kuo, J., Gandhi, J. G., Zia, R. N. & Paszek, M. J. Physical biology of the cancer cell glycocalyx. Nat. Phys. 14, 658–669 (2018).
Lakshminarayanan, A., Richard, M. & Davis, B. G. Studying glycobiology at the single-molecule level. Nat. Rev. Chem. 2, 148–159 (2018).
Almahayni, K. & Mockl, L. Setting the stage for universal pharmacological targeting of the glycocalyx. Curr. Top. Membr. 91, 61–88 (2023).
Dobie, C. & Skropeta, D. Insights into the role of sialylation in cancer progression and metastasis. Br. J. Cancer 124, 76–90 (2021).
Bordron, A. et al. Hyposialylation must be considered to develop future therapies in autoimmune diseases. Int. J. Mol. Sci. 22, 3402 (2021).
Unterauer, E. M. et al. Spatial proteomics in neurons at single-protein resolution. Cell 187, 1785–1800.e1716 (2024).
Jungmann, R. et al. Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT. Nat. Methods 11, 313–318 (2014).
Budiarta, M., Streit, M. & Beliu, G. Site-specific protein labeling strategies for super-resolution microscopy. Curr. Opin. Chem. Biol. 80, 102445 (2024).
Hudak, J. E., Canham, S. M. & Bertozzi, C. R. Glycocalyx engineering reveals a Siglec-based mechanism for NK cell immunoevasion. Nat. Chem. Biol. 10, 69–75 (2014).
Neelamegham, S. et al. Updates to the symbol nomenclature for glycans guidelines. Glycobiology 29, 620–624 (2019).
Strauss, S. & Jungmann, R. Up to 100-fold speed-up and multiplexing in optimized DNA-PAINT. Nat. Methods 17, 789–791 (2020).
Halpern, A. R., Howard, M. D. & Vaughan, J. C. Point by point: an introductory guide to sample preparation for single-molecule, super-resolution fluorescence microscopy. Curr. Protoc. Chem. Biol. 7, 103–120 (2015).
Edelstein, A. D. et al. Advanced methods of microscope control using muManager software. J. Biol. Methods 1, e10 (2014).
Schnitzbauer, J., Strauss, M. T., Schlichthaerle, T., Schueder, F. & Jungmann, R. Super-resolution microscopy with DNA-PAINT. Nat. Protoc. 12, 1198–1228 (2017).
Ma, H., Chen, M., Nguyen, P. & Liu, Y. Toward drift-free high-throughput nanoscopy through adaptive intersection maximization. Sci. Adv. 10, eadm7765 (2024).
Harris, C. R. et al. Array programming with NumPy. Nature 585, 357–362 (2020).
Virtanen, P. et al. SciPy 1.0: fundamental algorithms for scientific computing in Python. Nat. Methods 17, 261–272 (2020).
Bentley, J. L. Multidimensional binary search trees used for associative searching. Commun. ACM 18, 509–517 (1975).
Pettersen, E. F. et al. UCSF Chimera—a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).
Masullo, L. A. github.com/lumasullo/glycans-resi. GitHub (2025).
Ives, C. M. et al. Restoring protein glycosylation with GlycoShape. Nat. Methods 21, 2117–2127 (2024).
