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EMS Technical Data Sheets
PTAH Method for Central Nervous System Tissue
EMS Catalog #: 26658-Series
Fixation:
Formalin, 10% Neutral Buffered.
Sections:
Paraffin @ 6 microns
Staining Procedures:
- Deparaffinize and hydrate to distilled water.
- Incubate sections in Russell Modified Zenker’s Solution (#26658-05) (with Acetic Acid) at 60°C, 1 hour. Wash sections in running water, 15 minutes.
- Tone in Lugol’s Iodine (#26658-04) 15 minutes, then decolorize in 96% alcohol, 1 hour. Do not use Sodium Thiosulfate in this procedure because it will impair the further staining technique.
- Wash in distilled water for 5 minutes, and then oxidize with Potassium Permanganate Solution (#26658-01) for approx. 5 minutes.
- Again decolorize the section using Oxalic Acid, 5% (#26658-03), 5 minutes, then wash in distilled water followed by running water for 5 – 10 minutes.
- Stain in the PTAH (Phosphotungstic Acid – Hematoxylin) (#26658-02) reagent, 24 hours.
- Dip quickly into 95% alcohol, then further in absolute alcohol, two fast changes. Clear the slides in two changes of Xylene.
- Mount
Stain Results:
| Nuclei, Fibrin, Fibroglia, Microglia | Blue |
| Elastic Fibrils, Coarse | Purplish |
References:
AFIP Manual of Histologic Staining Technique; ed. G. Luna., 3rd ed. McGraw-Hill Publications, c. 1968, p. 199.