Staining solution reagent 30-30118LY

for histology

Product for the preparation of cyto-histological samples for optical microscopy. Application Cryostatic sections of 8 microns of human skeletal muscle. To show the enzymatic activity of acid phosphatase. Acid phosphatase is in macrophage and lysosome, it identifies necrosis and regeneration. Principle Results Naphthol acid phosphate is hydrolysed by acid phosphatases present in the tissue, and naphthol derivatives are produced. The naphthol derivatives couple with an unstable diazonium salt, hexazonium pararosaniline, to produce a red azo dye to mark the sites of enzyme activity. Method 1) It is recommend letting the reagents reach room temperature before starting the procedure. 2) Section in Fixative for acid phosphatase (Bio-Optica code 30-30120), 10 minutes 3) Dry in the air, 10 minutes 4) Preparation of incubating solution: - add reagent B to reagent C. Shake 5-10 seconds and allow to stand for 1 minute. - put the content of tube A into the reagent D. Shake 5-10 seconds. - add B+C solution to A+D solution. Shake 5-10 seconds. 5) Incubate sections at 37°C, 45 minutes. 6) Rinse in distilled water, 10 minutes 7) Methyl green (Bio-Optica code 30-30119), 5 minutes 8) Rinse in distilled water 9) Mount in aqueous mounting media. Alternatively dehydrate rapidly through alcohols to xylene and mount in resinous mounting media. Positive acid phosphatase enzymatic activity: red Results Background and nucleus: green