Staining solution reagent Von Kossa

for cytologyfor histology

Product for the preparation of cyto-histological samples for optical microscopy. To demonstrate deposits of calcium ions in tissue sections. PRINCIPLE This method is based on a substitution reaction. Tissue is treated with a silver nitrate solution; cationic silver replaces calcium in the original salt thus forming a silver salt that can be showed by a reduction to metallic silver. Reaction is as follows: CaCO3 + AgNO3 = Ag2CO3 + Ca(NO3)2 Ag2CO3 + 2H + = 2Ag + H2O + CO2 Calcium salts liable to react with silver nitrate are carbonate, phosphate, oxalate, sulphate, urate, chloride, sulfocyanide. Since calcium is present in mammalian tissue essentially in the form of calcium carbonate and calcium phosphate, this method is suitable to point it out. To prevent false results caused by the presence of uric acid and urates in the tissue, these salts have been solubilized in a lithium carbonate saturated solution. METHOD 1) Bring section to distilled water. 2) Put on the section 10 drops of reagent A: leave to act 10 minutes. 3) Rinse well in distilled water. 4) Put on the section 10 drops of reagent B: leave to act in the dark 1 hour. 5) Rinse well in distilled water. 6) Put on the section 10 drops of distilled water and add 10 drops of reagent C: leave to act 5 minutes (Until silver salts become black). 7) Rinse in distilled water. 8) Put on the section 10 drops of reagent D: leave to act 5 minutes. 9) Rinse in distilled water. 10) Put on the section 10 drops of reagent E: leave to act 5 minutes. 11) Rinse in distilled water and dehydrate through ascending alcohols, clear in xylene and mount.