AChE-Metachromatic Nissl Stain Procedure

Solutions

• 50 mM Sodium Acetate Buffer pH 5.0
• 1 litre distilled H₂O
• 6.8 g Sodiium Acetate
• 1.0 g Copper Sulphate (CuSO₄.5H₂O)
• 1.2 g Glycine
• Esterase Incubation Solution
• prepare this solution fresh and discard after use.
• 200 ml Sodium Acetate Buffer
• 232 mg S-acetylthiocholine iodide
• 6 mg ethoprpazine
• Acid-Acetone
• 100 ml Acetone
• 100 ml Glacial Acetic Acid
• 0.1% Cresyl Violet
• 1000 ml distilled H₂O
• 1 g Cresyl Violet
• Stir with spin bar several hours and filter
• 5% Acetic Acid
• 190 ml distilled H₂O
• 10 ml Glacial Acetic Acid
• 10% Potassium Ferricyanide
• 20 g Potassium Ferricyanide
• 200 ml distilled H₂O

Procedure

1. Esterase Incubation Solution .... 2 Hrs.-Overnight
2. Acid-Acetone.......................... 5 min.
3. 0.1% Cresyl Violet.................. 5 min.
4. 5% Acetic Acid....................…1 min.(or until differentiated)
5. 10% Ferricyanide ................... 2 min.
6. distilled H₂O ........................... 1 min.
7. acetone.................................. 10 sec
8. acetone.................................. 30 sec
9. Xylene..................................... 1 min.
10. Xylene..................................... 1 min.
11. coverslip

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Steve Milway

Last modified: Wednesday, December 17, 2003