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Mallory’s PTAH

Mallory's PTAH

6
steps
3
materials

Materials

MaterialAmountFunction
Hematein,1 g.Dye
Phosphotungstic acid10 gMordant
Distilled water1 LSolvent

Compounding Procedure

  1. Dissolve the hematein in 200 mL of the water and the phosphotungstic acid in the rest.
  2. Combine the solutions.
  3. Keep in a bottle with a tightly fitting cap to ensure atmospheric oxygen is excluded.
  4. The stain may be used after a day or two.

Protocol

  1. Bring sections to water with xylene and ethanol.
  2. Perform a Mallory bleach using 0.25% potassium permanganate.
  3. Rinse well with water.
  4. Place into the PTAH solution for 12 – 24 hours at room temperature.
  5. Rinse well with water.
  6. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei, erythrocytes, fresh fibrin, muscle striations  –  blue
  • Background  –  red

Notes

  • If formalin fixed, the sections may be treated with acid dichromate for 30 minutes.
    MaterialAmount
    Potassium dichromate, 3% aqueous36mL
    10% hydrochloric acid in absolute ethanol.12mL
  • The staining may be done in an oven at about 60°C for a few hours, but the results are often less satisfactory.
  • Many substances have been stated to be stained blue. Sometimes the red staining overshadows blue staining, and the section may need to be washed with water to remove excess red. Excess blue may be removed by extending the treatment with ethanol during dehydration.
  • PTAH may also be made with hematoxylin and oxidized as any other hematoxylin solution. Atmospheric oxidation is often recommended, but takes a few months. Place the solution in a flask with a loose cotton batting stopper to facilitate exposure to air. Test periodically and, when staining is satisfactory, place into a bottle with a tightly fitting lid to inhibit atmospheric oxidation.
  • Alternatively, the hematoxylin may be chemically oxidized. This is usually done with 12 mL of 1% aqueous potassium permanganate. The solution may be used after a day or two. Sodium iodate may also be used. Half the hematoxylin will be oxidized with 0.1 grams sodium iodate. If brought to a boil and allowed to cool, it may be used immediately, or it may be allowed to oxidize at room temperature for a few days.

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.

References

  1. Bancroft, J. D. and Stevens, A.,
    Theory and Practice of Histological Techniques, Ed. 2,
    Churchill Livingstone, London, UK.