Histotechnology Technical methods

STAIN FOR VIRAL INCLUSIONS

Methods Index

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PHLOXINE TARTRAZINE

Solutions

a) Phloxine solution

--Phloxine 0.5g

--Calcium chloride 0.5g

--Distilled water 100ml

b) Tartrazine solution

--Saturated solution of tartrazine in 2-ethoxyethanol ('cellosolve').


Method

1. Dewax in xylene, then hydrate through graded alcohols to water.

2. Stain nuclei in alum haematoxylin (e.g. Carazzi's for 10 minutes).

3. Wash in running tap water for 5 minutes.

4. Stain in phloxine solution for 20 minutes.

5. Rinse in tap water, then blot dry.

6. Stain in tartrazine solution under microscopic control until only the viral inclusions remain strongly red. An average time is 5-10 minutes.

7.Rinse in 95% alcohol.

8. Dehydrate in absolute alcohol, clear . Mount sections in DPX


RESULTS

--Viral inclusions - bright red

--Red blood cells - variably orange-red

--Nuclei - blue-grey

--Background - yellow

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