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Histology Lab: E.P.S.R.
Histotechnology Technical methods

Connective Tissue Stain

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E.P.S.R.

Elastic Picro Sirius Red

1. Bring sections to water

2. Acidified potassium permanganate 2 minutes 3. Wash in water

4. 1% Oxalic acid 1 minute

5. Wash in water

6. Wash in 70% alcohol (optional)

7. Stain in Weigert's Resorcin Fuchsin at room temperature 45 minutes

8. Wash in water 10 minutes

9. Differentiate in Acid alcohol

10. Wash in water 10 minutes

1. Stain in Picro-sirius red 1 hour

12. Wash in water

13. Dehydrate, clear . Mount sections in DPX

N.B. Wash quickly at step 12. Dehydrate quickly


RESULTS:

--Collagen - Red

--Nuclei - Blue-Black

--Background - Yellow

--Elastin Black


SOLUTIONS:

Picro-Sirius red 0.1 Sirius red F3B in saturated Aqueous Picric Acid

Weigert Resorcin Fuchsin

1. To 100ml of distilled water Add 1g of basic fuchsin and 2g of resorcin.

2. Boil

3. Add 12.5ml of freshly prepared 30% Ferric Chloride solution

4. Continue boiling for 5 minutes

5. Cool and filter, discarding the filtrate

6. Dissolve the precipitate in 100ml of 95% Ethanol using a hot plate or water bath for controlled heating.

7. Add 2ml of concentrated HCl

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