Histotechnology Technical methods

Stain for Reticulin

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GORDON AND SWEETS RETICULIN METHOD

SOLUTIONS

1 Silver Solution

To 5cm3 of 10% silver nitrate, add concentrated ammonia drop by drop until the precipitate has just dissolved. Add 5cm3 of 3% sodium hydroxide and redissolve the precipitate formed with a few drops of ammonia. Dilute to 50cm3 with distilled water.

2. 1% Acidified Potassium Permanganate

3. 1% Oxalic Acid

4. 2.5% Iron Alum

5. 10% Aqueous Formalin

6. 0.5% Cold Chloride

7. 5% Sodium Thiosulphate ("HYPO")

8. 1% Eosin


METHOD:

1. Sections to water.

2. Treat slides with 1% acidified potassium permanganate for 2 minutes.

3. Rinse in distilled water.

4. Bleach in 1% oxalic acid solution.

5. Rinse in distilled water.

6. Treat slides with 2.5% iron alum for 10 minutes.

7. Wash well in several changes of distilled water.

8. Treat slides with silver solution (kept at 4ÉC) for 20 seconds.

9. Wash well in several changes of distilled water.

10. Reduce in 10% aqueous formalin solution till the sections turn black.

11. Rinse in tap water.

12. Tone in 0.5% gold chloride for 1 minute if desired.

13. Rinse in tap water.

14. Treat slides with 5% sodium thiosulphate for 5 minutes.

15. Rinse in tap water.

16. Counterstain in eosin for 1 minute.

17. Dehydrate, clear . Mount sections in DPX


RESULTS:

--Reticulin fibres - black

--Nuclei - black or unstained.

--Other elements according to the counterstain.

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.


© 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