Use of semithin sections for the histochemical study of carbohydrate-containing biopolymers in cells and tissues with the aid of lectins
Lutsik, A.D.; Kotyk, A.E.
Biulleten' Eksperimental'noi Biologii i Meditsiny 100(12): 755-758
1985
ISSN/ISBN: 0365-9615 PMID: 3907731 Document Number: 248095
A sensitive method of histochemical analysis of cellular and tissue glycoconjugates on semithin sections using lectins is suggested. For fixation tissue bioptates were incubated for 4 h in a 2.5% glutaraldehyde in phosphate buffered saline (PBS) at 4 degrees C, then washed for 1 h in 0.2 M glycine in PBS. After epon-araldite embedment and preparation of semithin sections, the resin was removed in saturated ethanol-KOH solution during 5-10 s. Endogenous perooxidase was inactivated in methanol containing 0.3% H2O2. For identification of lectin-binding sites semithin sections were incubated for 30 min in a 0.005% solution of lectin-peroxidase conjugate in PBS and visualized by 0.05% diaminobezidine solution in PBS, containing 0.015% H2O2. The method described ensures good preservation of cellular and tissue glycoconjugates and is highly specific and sensitive.