Histochemistry

, Volume 89, Issue 3, pp 277–282 | Cite as

Light and electron microscopical postembedding lectin histochemistry for WGA-binding sites in the renal cortex of the mouse embedded in polyhydroxy aromatic resins LR-White and LR-Gold

  • R. Herken
  • M. Fussek
  • M. Thies
Article

Summary

This work describes a technique which permits study of the postembedding lectin histochemistry for WGA-binding sites at the light and electron microscopical level on the same resin embedded tissue without removing or etching of the resin. Unfixed kidney pieces or kidney pieces fixed in 4% formaldehyde were embedded in the hydrophilic polyhydroxy aromatic resins LR-Gold and LR-White, following dehydration in up to 70% ethanol, 90% ethanol or 100% ethanol. LR-Gold was cryopolymerised at −25° C using the light sensitive initiator benzil, whereas LR-White was heat-cured at +50° C. The localisation of WGA-binding sites at the light microscopical level was investigated using FITC-labelled WGA. The ultrastructural localisation of WGA-binding sites was performed using 15 nm gold-labelled WGA. The best fluorescence staining results were obtained on fixed or unfixed tissue dehydrated in up to 70% ethanol and embedded in LR-Gold. At the ultrastructural level, the best staining results for WGA-binding sites were seen on tissue sections, dehydrated in up to 90% ethanol prior to embedding in LR-Gold.

Keywords

Formaldehyde Tissue Section Microscopical Level Renal Cortex Fluorescence Staining 

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References

  1. Bretton R, Bariety J (1976) A comparative ultrastructural localisation of concanavalin A, wheat germ and ricinus communis on glomeruli of normal rat kidney. J Histochem Cytochem 24:1093–1100Google Scholar
  2. Ellinger A, Pavelka M (1985) Post-embedding localisation of glycoconjugates by means of lectins on thin sections of tissue embedded in LR-White. Histochem J 17:1321–1335Google Scholar
  3. Farnum CE, Wilsman NJ (1984) Lectin-binding histochemistry of non-decalcified growth plate cartilage: A postembedment method for light microscopy of Epon-embedded tissue. J Histochem Cytochem 32:593–607Google Scholar
  4. Holthöfer H (1983) Lectin binding sites in kidney: A comparative study of 14 animal species. J Histochem Cytochem 31:531–537Google Scholar
  5. Holthöfer H, Virtanen I, Pettersson E, Törnroth T, Alfthan O, Linder E, Miettinen A (1981) Lectins as fluorescence microscopic markers for saccharides in the human kidney. Lab Invest 45:391–399Google Scholar
  6. Jones CJP, Stoddard RW (1986) A postembedding avidin-biotin peroxidase system to demonstrate the light and electron microscopical localization of lectin binding sites in rat kidney tubules. Histochem J 18:371–379Google Scholar
  7. Lucocq JM, Roth J (1984) Demonstration of antigenic and lectinbinding sites in semithin resin sections. J Histochem Cytochem 32:1075–1083Google Scholar
  8. Newman GR (1987) Use and abuse of LR-White. Histochem J 19:118–120Google Scholar
  9. Newman GR, Jasani B, Williams ED (1983) A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope. Histochem J 15:543–555Google Scholar
  10. Roth J (1983) Application of immunocolloids in light microscopy. Demonstration of lectin-binding sites in paraffin sections by the use of lectin-gold or glycoprotein-gold complexes. J Histochem Cytochem 31:547–552Google Scholar
  11. Roth J, Brown D, Orci L (1983) Regional distribution of N-acetyl-d-galactosamine residues in the glycocalyx of glomerular podocytes. J Cell Biol 96:1189–1196Google Scholar
  12. Roth J, Lucocq M, Charest PM (1984) Light and electron microscopic demonstration of sialic acid residues with the lectin from limax flavus: A cytochemical affinity technique with the use of fetuin-gold complexes. J Histochem Cytochem 32:1167–1179Google Scholar
  13. Yokota S, Oda T (1985) Immunocytochemical demonstration of serine: Pyruvate aminotransferase in peroxisomes and mitochondria of rat kidney. Histochemistry 83:81–85Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • R. Herken
    • 1
  • M. Fussek
    • 1
  • M. Thies
    • 1
  1. 1.Abteilung HistologieZentrum AnatomieGöttingenFederal Republic of Germany

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