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.
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.
D000180:Acrylic Resins; D000818:Animals; D001665:Binding Sites; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006651:Histocytochemistry; D007672:Kidney Cortex / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008854:Microscopy, Electron; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins / Q000378:metabolism*
D000180:Acrylic Resins; D005452:Fluoresceins; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins; C048707:LR white; C056756:LR gold; D016650:Fluorescein-5-isothiocyanate