Please find the following media preparation method:
New Fuchsin Alkaline Phosphatase Substrate Solution Red
0.01% New Fuchsin, 0.02% Sodium Nitrite, 0.05%
Naphthol AS-BI Phosphate, 1mM Levamisole, in 0.05M
Tris-HCl, pH 8.7
0.2% New Fuchsin (20x) in 2N HCl and store at 4 °C.
0.4% Sodium Nitrite (20x) and 20mM Levamisole (20x) in
distilled water. Store at 4 °C.
1% Naphthol AS-BI Phosphate (20x) (Sigma N2250) in
100% DMF (dimethylformamide) and store at 4 °C.
To 5 ml of 0.05M Tris-HCl, pH8.7,
Add 5 drops of 0.2% New Fuchsin and mix well.
Add 5 drops of 0.4% Sodium Nitrite/20mM Levamisole and
Add 5 drops 1% Naphthol AS-BI Phosphate and mix well.
Incubate sections for 10-20 minutes at room temperature.
Rinse in tap water, counterstain and coverslip with
permanent mounting medium.