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.

Abstract

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 degrees C using the light sensitive initiator benzil, whereas LR-White was heat-cured at +50 degrees 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.

• Authors: R Herken, M Fussek, M Thies
• Journal: Histochemistry (Histochemistry) Vol. 89 Issue 3 Pg. 277-82 ( 1988) ISSN: 0301-5564 [Print] Germany
• PMID: 3136102 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Acrylic Resins
• Fluoresceins
• Thiocyanates
• Wheat Germ Agglutinins
• LR white
• LR gold
• Fluorescein-5-isothiocyanate

Topics

• Acrylic Resins
• Animals
• Binding Sites
• Female
• Fluorescein-5-isothiocyanate
• Fluoresceins
• Histocytochemistry
• Kidney Cortex (metabolism, ultrastructure)
• Mice
• Microscopy, Electron
• Thiocyanates
• Wheat Germ Agglutinins (metabolism)