PERIODIC ACID, SCHIFFS

1. Bring sections to water.

2. Treat with 1% periodic acid for 10 mins.

3. Wash in water.

4. Treat with Schiffs reagent for 10 mins.

5. Wash well in tap water.

6. Stain nuclei with Carazzi haematoxylin for 2 mins.

7. Differentiate in acid alcohol.

8. Blue in Scotts tap water.

9. Dehydrate, clear . Mount sections in DPX

N.B. Do not over stain in schiffs reagent - it is irreversible. Wash for several mins at step 5 to bring out the colour of the Schiffs.

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RESULTS

--Glycogen, other periodate reactive carbohydrates - magenta

--Nuclei- blue

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SOLUTIONS

Schiffs reagent

1. Boil 200ml of distilled water, remove flask from bunsen and add 1g of basic fuchsin.

2. Allow to cool to 50°c.

3. Add 2g of potassium metabisulphate whilst mixing.

4. Cool to room temperature and add 2ml of concentrated hydrochloric acid mix and stand in the dark overnight.

5. Add a large amount of activated charcoal, shake well and filter. The solution should be clear/pale yellow.

Store at 4°c

NOTE: No responsibility is assumed by The University of Nottingham or the Queens Medical Centre NHS Trust for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. It is the users responsibilty to ensure that all procedures are carried out according to appropriate Health and Safety requirements.

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© Copyright 1997 University of Nottingham Medical School Division of Histopathology. This page was last built on Mon, May 5, 1997 with Frontier. Thanks for looking in. Comments to James.Lowe@nottingham.ac.uk