[Histonet] PAS PROCEDURE FROM MUSCLE/NERVE PACKET

From:"George Cole"

For Those Who Received Muscle & Nerve packets;

Sorry, when I made up the packets, I used forms filled out for our last inspection by office staff.  The typing was so much better than my own.  All the ingredients were listed, but the actual PAS procedure was left out.

The following procedure is done after the PAS reagents are prepared:

 

This follows page 12:

 

1. Put slides in the fixative    10 minutes to one hour.

2. Rinse well in deionized water.

3. Place in freshly mixed  0.5% aqueous Periodic Acid     5 minutes.

4. Wash thoroughly in deionized water.

5. Place in Schiff’s Reagent      15 minutes.

6. Rinse in three changes of acid rinse       for two minutes each.

7.  Wash in gently running water     for a minute or two.  Rinse with deionized water.  Stain with Gill’s III  hematoxylin for

 30 seconds to a minute.  Blue and rinse with running tap water   15 minutes.   Rinse in deionized water.

8 . Dehydrate with 95% and 100% alcohols.  Clear in xylene and mount in Permount or equivalent.

 

DIASTASE  DIGESTION  PROCEDURE:

 

1. Fix for 10 minutes.

2. Rinse well.

3. Digest in filtered Diastase solution for one hour at 37C.

4.  Rinse well in deionized water.

5 Proceed with regular PAS routine from step 3 above.

 

     


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