ALKALINE PHOSPHATASE

SECTIONS

Fresh Frozen (jejunal Biopsy) or cold Formal calcium/Gum Sucrose fixed Cryostat 5-7micron thick.
 

 

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SOLUTIONS

a. 0.2M Tris HCL buffer pH 8.3

• 0.2M Tris (2.42g/100) 25ml 125ml )
• 0.1N HCL (0.85/110ml) 20ml 100ml ) 500ml
• Distilled water 55ml 275ml )

b. Naphthol ASBI stock

• Naphthol ASBI phosphate 25mg
• N:N Dimethyl formamide 10ml
• Distilled water 10ml
• Adjust to pH 8.0 with 1M sodium carbonate if necessary then add 300 ml distilled water
• 0.2M Tris HCL buffer pH 8.3 (solution a.) 180ml

c. Incubating solution

• Naphthol ASBI stock (solution b.) 10ml
• Fast red TR 10mg
• Filter, use immediately

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METHOD

 

1. Incubate in solution c. for 5-15 minutes at room temperature.
2. Wash, counterstain in haematoxylin, wash.
3. Mount in phosphate buffered glycerine jelly

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RESULTS

 

• Alkaline phosphatase activity - Red
• Nuclei - Blue