Skip to main content
SpringerLink
Log in

Lectin—Gold Histochemistry on Paraffin and Lowicryl K4M Sections Using Biotin and Digoxigenin-Conjugated Lectins

  • Protocol
Lectin Methods and Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 9))

  • 1810 Accesses

Abstract

A variety of staining reactions for the visualization of cellular and extracellular glycoconjugates at the light microscopic level are available that are based on the detection of carboxyl and sulfate groups or periodic acid reactive configurations (1,2). Starting in the late 1960s lectins have replaced many of these chemical staining reactions because of their high specificity for defined monoand oligosaccharidic sequences in both N- and o-glycosidic-linked oligosaccharide side-chains of glycoproteins and glycolipids. In order to be useful as histochemical reagents, lectins must be tagged with appropriate markers and those employed in immunocytochemistry have been used successfully (3-9) Horisberger and coworkers were the first to prepare lectins labeled with particles of colloidal gold and used them in scanning electron mrcroscopy (10) Subsequently, gold-labeled lectins were applied in transmission electron microscopy to study various aspects of cell surface expression and internalization of lectin-binding sites (5,8,11,12), as well as in postembedding labeling of Lowicryl K4M thin sections (13). Later, it was shown that gold-labeled lectins can be used to stain sections of paraffin-embedded tissues (14-16), as well as semithin sections of Epon (17) and Lowicryl K4M-embedded tissues (18,19), However, in order to achieve a visible pink staining, which is the natural color of particles of colloidal gold in transmitted visible light (20), highly concentrated lectin-gold complexes had to be used, thereby allowing the possibility of nonspecific staining. A major improvement resulted through the application of a photochemrcal silver reaction for signal amphfication (21-24), whrch permitted the use of lectins for light microscopy in concentrations as applied for electron microscopy (25,26).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Spicer, S. S., Baron, D S, Sato, A., and Schulte, B. A. (1981) Variability of cell surface glycoconjugates. Relation to differences in cell function. J Histochem Cylochem 29,994

    CAS  Google Scholar 

  2. Reid, P E and Park, C M. (1990) Carbohydrate Histochemistry of Epithelial Glycoproteins Gustav Fischer Verlag, Stuttgart, pp 1–170.

    Google Scholar 

  3. Nicolson, G. (1974) The interaction of lectins with animal surfaces Int Rev Cytol 39, 89–190

    Article  PubMed  CAS  Google Scholar 

  4. Roth, J (1978) The lectins Molecular probes in cell biology and membrane research Exp Path Suppl 3, 1–186

    Google Scholar 

  5. Roth, J. and Binder, M (1978) Colloidal gold, ferritin and peroxidase as markers for electron microscopic double labeling lectin techniques J Histochem Cytochem 26, 163–169

    PubMed  CAS  Google Scholar 

  6. Roth, J and Thoss, K (1974) Light and electron microscopic demonstration of D-mannose and D-glucose like sites at the cell surface by means of the lectin from the Lens culinaris Experientra 3, 414

    Article  Google Scholar 

  7. Roth, J and Thoss, K (1975) The use of fluorescein isothiocyanate labeled lectins for immunohistochemical demonstration of saccharides 1 Methodical investigations with Concanavalin A, Lens culinaris lectin and Ricinus communis lectin Exp Path 10,258–267

    CAS  Google Scholar 

  8. Roth, J and Wagner, M. (1977) Peroxidase and gold complexes of lectins for double labeling of cell surface binding sites by electron microscopy J Hitochem Cytochem 25, 1181–1184

    CAS  Google Scholar 

  9. Roth, J, Binder, M, and Gerhard, J (1978) Conjugation of lectins with fluorochromes. an approach to histochemical double labeling of carbohydrate components. Histochemistry 56, 265–273

    Article  PubMed  CAS  Google Scholar 

  10. Horisberger, M, Rosset, J, and Bauer, H. (1975) Colloidal gold granules as markers for cell surface receptors in the scanning electron microscope. Experientia 31, 1147

    Article  PubMed  CAS  Google Scholar 

  11. Horisberger, M. and Rosset, J (1977) Colloidal gold, a useful marker for transmission and scanning electron microscopy. J Histochem Cytochem 25, 295–305

    PubMed  CAS  Google Scholar 

  12. Wagner, M, Roth, J, and Wagner, B (1976) Gold-labeled protectin from Helix pomatra for the localization of blood group A antigen from human erythrocytes by immuno freeze etching Exp Path 12,277–281

    Google Scholar 

  13. Roth, J (1983) Application of lectin-gold complexes for electron microscopic localization of glycoconjugates on thin sections J Histochem Cytochem 31, 987–999

    PubMed  CAS  Google Scholar 

  14. Roth, J (1983) Application of immunocolloids in light microscopy. II Demonstration of lectin-gold complexes or glycoprotein-gold complexes J Histochem Cytochem 31,547–552

    PubMed  CAS  Google Scholar 

  15. Roth, J. (1983) in Techniques in Immunocytochemistry (Bullock, G and Petrusz, P, eds), Academic, London, pp. 217–284

    Google Scholar 

  16. Lucocq, J and Roth, J (1985) in Techniques in Immunocytochemistry (Bullock, G and Petrusz, P, eds), Academic, London, pp. 203–236

    Google Scholar 

  17. Lucocq, J and Roth, J (1984) Applications of Immunocolloids in light microscopy. III. Demonstration of antigenic and lectin-binding sites in semithin resin sections J Histochem Cytochem 32, 1075–1083

    PubMed  CAS  Google Scholar 

  18. Roth, J., Brown, D, and Orci, L. (1983) Regional distribution of N-acetyl-Dgalactosamine residues in the glycocalyx of glomerular podocytes J Cell Biol 96,1189–1196

    Article  PubMed  CAS  Google Scholar 

  19. Roth, J (1989) in Vesicular transport Methods in Cell Biology (Tartakoff, A, ed.), Academic, San Diego, pp 513–551

    Google Scholar 

  20. Faraday, M (1857) Experimental relations of gold (and other metals) to light Phil Trans R Soc 147, 145–181

    Article  Google Scholar 

  21. Danscher, G (1981) Localisation of gold in biological tissue. A photochemical method for light and electron microscopy Histochemistry 71, 81–88

    Article  PubMed  CAS  Google Scholar 

  22. Danscher, G and Rytter Norgaard, J O (1983) Light microscopic visualization of colloidal gold on resin-embedded tissue J Histochem Cytochem 31, 1394–1398

    PubMed  CAS  Google Scholar 

  23. Holgate, C S, Jackson, P, Cowen, P N, and Bird, C C (1983) Immunogoldsilver staining new method of Immunostaining with enhanced sensitivity J Histochem Cytochem 31, 938–944

    PubMed  CAS  Google Scholar 

  24. Springall, D R, Hacker, G W, Grimelius, L, and Polak, J M (1984) The potential of the Immunogoldsilver staining method for paraffin sections Histochemistry 81, 603–608

    Article  PubMed  CAS  Google Scholar 

  25. Taatjes, D J, Schaub, U, and Roth, J (1987) Light microscopical detection of antigens and lectin binding sites with gold labeled reagents on semithin Lowicryl K4M sections usefulness of the photochemical silver reaction for signal amplfication Histochem J 19, 235–245

    Article  PubMed  CAS  Google Scholar 

  26. Taatjes, D J, Roth, J, Peumans, W, and Goldstein, I J (1988) Elderberry bark lectin-gold techniques for the detection of Neu5Ac(α2,6)GalαlNAc sequences applications and limitations Histochem J 20, 478–490

    Article  PubMed  CAS  Google Scholar 

  27. Taatjes, D and Roth, J (1991) Glycosylation in intestinal epithelium Int Rev Cytol 126, 135–193

    Article  PubMed  CAS  Google Scholar 

  28. Carlemalm, E., Garavito, R M, and Villiger, W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature J Microsc (Oxford) 126, 123–143

    CAS  Google Scholar 

  29. Roth, J, Bendayan, E., Carlemalm, E, Villiger, W, and Garavito, M (1981) The enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue J Histochem Cytochem 29, 663–671

    PubMed  CAS  Google Scholar 

  30. Haselbeck, A., Schickaneder, E, and Hosel, W (1990) Structural characterization of glycoprotein carbohydrate chains by using digoxigenin-labelled lectins on blots Anal Biochem 191, 25–30

    Article  PubMed  CAS  Google Scholar 

  31. Haselbeck, A., Schikander, E., Schmidt, A., v d Eliz, H, and Hosel, W (1989) New Techniques for the ldentification and characterization of glycoproteins on gels and blots, Proc 19th FEBS meeting, Rome

    Google Scholar 

  32. Sata, T., Zuber, C, and Roth, J (1990) Lectin digoxigenin conjugates. a new hapten system for glycoconjugate cytochemistry Histochemistry 94, 1–11

    Article  PubMed  CAS  Google Scholar 

  33. Wood, G and Warnke, R (1981) Suppression of endogenous avidin-binding activity in tissues and its relevance to biotin-avidin detection systems J Histochem Cytochem 29, 1196–1204

    PubMed  CAS  Google Scholar 

  34. Skutelsky, E, Goyal, V, and Alroy, J (1987) The use of avidin-gold complex for light microscopic localization of lectin receptors Histochemistry 86, 291

    Article  PubMed  CAS  Google Scholar 

  35. Hacker, G W, Grimelius, L, Danscher, G, Bernatzky, G, Muss, W, Adam, H, and Thurner, J (1988) Silver acetate autometallography. an alternative enhancement technique for immunogold-sliver staining (IGSS) and silver amplification of gold, silver, mercury and zinc in tissues J Histotechnol 11, 213–221

    CAS  Google Scholar 

  36. Sata, T, Lackie, P, Taatjes, D, Peumans, W, and Roth, J (1989) Detection of the NeuAc (α2,3) Gal (β1,4) GlcNAc sequence with the leukoagglutinin from Maackia amurensis light and electron microscopic demonstration of differential tissue expression of terminal sialic acid in α2,3 and α2,6-linkage J Histochem Cytochem 37, 1577–1588

    PubMed  CAS  Google Scholar 

  37. Sata, T, Zuber, C, Rinderle, S J, Goldstein, I J and Roth, J (1990) Expression patterns of the T antigen and the cryptic T antigen in rat fetuses detection with the lectin Amaranthin J Histochem Cytochem 38, 763–774

    PubMed  CAS  Google Scholar 

  38. Sata, T, Roth, R, Zuber, C., Stamm, B, and Heitz, P (1991) Expression of α2,6-linked sialic acid residues in neoplastic but not in normal colonic mucosa Am J Pathol 139, 1435–1448

    PubMed  CAS  Google Scholar 

  39. Taatjes, D and Roth, J (1990) Selective loss of sialic acid from rat small lntestinal epithelial cells during postnatal development demonstration with lectin-gold techniques Eur J Cell Biol 53, 255–266

    PubMed  CAS  Google Scholar 

  40. Schulte, B A and Spicer, S S (1983) Light microscopic histochemical detection of terminal galactose and N-acetylgalactosamine residues rodent complex carbohydrates using a galactose oxidase-Schiff sequence and the peanut lectin-horseradish peroxidase conjugate J Histochem Cytochem 31, 19–24

    PubMed  CAS  Google Scholar 

  41. Ravindranath, M. H., Higa, H. H, Cooper, E L, and Paulson, J C. (1985) Purification and characterization of an o-acetylsialic acid-specific from a marine crab Cancer antennarius J Blol Chem 260. 8850–8856

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Cell and Molecular Pathology, Department of Pathology, University of Zurich, Switzerland

    Jürgen Roth, Christian Zuber, Tetsutaro Sata & Wei-Ping Li

Authors
  1. Jürgen Roth
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christian Zuber
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tetsutaro Sata
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wei-Ping Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Liverpool, UK

    Jonathan M. Rhodes  & Jeremy D. Milton  & 

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc.

About this protocol

Cite this protocol

Roth, J., Zuber, C., Sata, T., Li, WP. (1998). Lectin—Gold Histochemistry on Paraffin and Lowicryl K4M Sections Using Biotin and Digoxigenin-Conjugated Lectins. In: Rhodes, J.M., Milton, J.D. (eds) Lectin Methods and Protocols. Methods in Molecular Medicine™, vol 9. Humana Press. https://doi.org/10.1385/0-89603-396-1:41

Download citation

  • DOI: https://doi.org/10.1385/0-89603-396-1:41

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-396-2

  • Online ISBN: 978-1-59259-593-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation