PHOSPHOTUNGSTIC ACID HAEMATOXYLIN

                    PHOSPHOTUNGSTIC ACID HAEMATOXYLIN

Solution
                   Haematoxylin                                                -       0.5gm
                   Phosphotungstic acid                                    -        10 gm
                   Distilled water                                               -        500 ml
                   25% Aqueous potassium permanganate         -        25ml.

The haematoxylin is dissolved in 100 ml of the distilled water and the phosphotungstic acid in the remaining 400 ml. The two solutions are mixed and the potassium rermanganate solution is added. The stain can be used next day, but peak staining activity is reached until after 7 days.

Procedure
1) Deparaffinise the section in xylene for 10 mts
2) Bring the section to water through descending series of alcohol.
3) Treat with 0.25% potassium permanganate for 5 mts.
4) Wash in water.
5) Treat with 5% oxalic acid for 2 mts
6) Rinse in distilled water.
7) Stain with PTAH solution for 16-18 hrs. 8) Dehydrate rapidly through 95% and absolute alcohol. 9) Clear and mount.

Result

Nuclei, fibrin, muscle striations, RBCS, neuroglia             -           Blue
Collagen, bone                                                                      -           Brick red or paler shade.


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