PTAH Method for Central Nervous System Tissue

EMS26658

Components:

Potassium Permanganate 1%

Phosphotungstic Acid-Haematoxylin

Oxalic Acid 5% Aqueous

Lugol's Iodine Solution

Russels Modified Zenkers Fluid

Fixation:

Formalin, 10% Neutral Buffered.

Sections:

Paraffin @ 6µm

Staining Procedures:            

1. Deparaffinise and hydrate to distilled water.
2. Incubate sections in Russell Modified Zenker’s Solution (with Acetic Acid) at 60°C, 1 hour. Wash sections in running water, 15 minutes.
3. Tone in Lugol’s Iodine15 minutes, then decolourise in 96% alcohol, 1 hour. Do not use Sodium Thiosulphate in this procedure because it will impair the further staining technique.
4. Wash in distilled water for 5 minutes, and then oxidise with Potassium Permanganate Solution for approx. 5 minutes.
5. Again decolourise the section using Oxalic Acid, 5%, 5 minutes, then wash in distilled water followed by running water for 5 – 10 minutes.
6. Stain in the PTAH (Phosphotungstic Acid – Haematoxylin) reagent, 24 hours.
7. Dip quickly into 95% alcohol, then further in absolute alcohol, two fast changes. Clear the slides in two changes of Xylene.
8.  Mount

Stain Results:  

References:

AFIP Manual of Histologic Staining Technique; ed. G. Luna., 3rd ed. McGraw-Hill Publications, c. 1968, p. 199.