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Electron Microscopy Sciences

Technical Data Sheets

Periodic Acid Schiff Stain For Glycol Methacrylate Sections

EMS Catalog #14653-14654

Technovit H7100 / H8100 Periodic Acid Schiff Stain

Procedure

  1. Oxidize sections in 0.4% periodic acid for 30 minutes at 57°C.
  2. Wash in running tap water or sodium sulfite solution (to avoid a general pink background).
  3. Rinse in three times in distilled water.
  4. Schiff's Reagent for 15 minutes.
  5. Wash well in running tap water.
  6. Rinse in distilled water.
  7. Counterstain with Gill's Hematoxylin for 10 minutes.
  8. Wash well in running tap water.
  9. Dehydrate using 95% and 100% ethanol.
  10. Clear in xylene and cover slip.

Results

Nuclei
blue
Glycogen
violet / red
Basement membranes
violet / red
Mucine
violet / red

Solutions

Gill's Hematoxylin:

  • Hematoxylin (C.I. 75290) - 6 gm
  • Sodium Iodate - 0.6 g
  • Aluminum Sulfate - 52.8 g
  • Distilled water - 690 ml
  • Ethylene Glycol - 250 ml
  • Glacial acetic acid - 60 ml

Schiff's Reagent :

  • Mix 0.5 g of Pararosaniline (C.I.#42500) into 15 ml of 1N Hydrochloric Acid.
  • Mix 0.5 g of Potassium Metabisulfite into 85 ml of distilled water.
  • Add the second solution to the first and store for 24 hours in a dark place.
  • Bleach the resulting solution with 200 mg of bone charcoal and filter.
  • Store the colorless solution at 4°C.

Sodium Sulfite Solution:

  • 0.5 g of sodium sulfite
  • 100 ml of distilled water

Product Information

Technovit Glycol Methacylate Kits H7100 / H8100