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Electron Microscopy Sciences

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:            

  1. Deparaffinize and hydrate to distilled water.
  2. 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.
  3. 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.
  4. Wash in distilled water for 5 minutes, and then oxidize with Potassium Permanganate Solution (#26658-01) for approx. 5 minutes.
  5. 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.
  6. Stain in the PTAH (Phosphotungstic Acid – Hematoxylin) (#26658-02) 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:  

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.

Online Ordering

PTAH Method for Central Nervous System Tissue is available online from the EMS Catalog. For ordering or product information, click here.