Histotechnology Technical methods

Stain for elastic

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8 µm paraffin wax sections.


1. 1% Acidified Potassium permanganate.

2. 1% Oxalic Acid.

3. Staining Solution.

  • Victoria Blue 4R 1g
  • New Fuchsin 1g
  • Crystal Violet 1g
Dissolve in 200ml of hot water, then add in the following order:-
  • Resorcin 4g
  • Dextrin 4g
  • 30% ferric chloride (freshly prepared) 50ml
Boil for 5 minutes and filter whilst hot. Transfer precipitate and filter paper to original beaker and redissolve in 200ml 95% alcohol. Boil for 15-20 minutes, filter and make up to 200ml with 95% alcohol. Finally add 2ml of concentrated hydrochloric acid.


  1. Sections to water.
  2. Treat with 1% acidified potassium permanganate for 2 minutes.
  3. Rinse in tap water.
  4. Bleach in 1% oxalic acid for 1 minute or until colourless.
  5. Wash in water, then in 95% alcohol.
  6. Put sections in staining solution for 1 hour.
  7. Wash in 95% alcohol.
  8. Counterstain as required in Van Gieson or picro-sirius red for 5 minutes.
  9. Dehydrate, clear . Mount sections in DPX


  • Elastic fibres and most cell granules - blue/black
  • Background - according to 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