Periodic Acid-Schiff’s Reagent (PAS Stain)

Reagent Preparation

Reagents are supplied ready for use. Bring Periodic Acid and Schiff’s Reagent to room temperature before use.

Formalin-Ethanol Fixative (Films)

Mix 5 mL of formaldehyde with 45 ml of 95% ethanol. Prepare fresh.

Procedure for Tissue Sections

  1. Deparaffinize tissue sections and hydrate to deionized water.
  2. Place in Periodic Acid solution for 5 minutes.
  3. Rinse in deionized water.
  4. Immerse in Schiff’s Reagent for 15 minutes.
  5. Wash in running tap water for 5 minutes.
  6. Counterstain in Gill’s Hematoxylin No. 3 for 90 seconds.
  7. Wash in running tap water.
  8. Dehydrate in alcohol, clear in xylene and mount.

Blood, Bone Marrow, or Touch Preps

  1. Fix air dried films in Formalin – Ethanol fixative for 1 minute.
  2. Rinse for 1 minute in running tap water.
  3. Place in Periodic Acid solution for 5 minutes.
  4. Rinse in several changes of deionized water.
  5. Immerse in Schiff’s Reagent for 15 minutes.
  6. Wash in running tap water for 5 minutes.
  7. Counterstain in Gill’s Hematoxylin No. 3 for 90 seconds.
  8. Wash in running tap water for 15-30 seconds and air dry.
  9. Examine under oil immersion or mount in xylene based mounting media.

Results

PAS positive substances will stain rose to magenta with blue to black nuclei.

Periodic Acid-Schiff's Reagent (PAS Stain)

Reagents

     
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