Histotechnology Technical methods

Stain for fine cell detail

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4µm paraffin wax sections


1. 0.25% Acidified Potassium Permanganate

2. 5% Oxalic Acid

3. P.T.A.H. --Haematoxylin 0.5g

--Phosphotungstic acid 10.0g

--Distilled water 500cm3

--0.25% aqueous potassium permanganate 25cm3

The haematoxylin is dissolved in 100cm3 of the distilled water, and the phosphotungstic acid in the remaining 400cm3; the two solutions are mixed and the potassium permanganate solution is added. The stain can be used the next day but peak staining activity is not reached until after 7 days. Continuing oxidation of the haematoxylin means the stain has comparatively short life.


1. Take sections to water

2. Treat sections with 0.25% acidified potassium permanganate for 5 minutes

3. Rinse in water and decolourise in 5% oxalic acid

4. Rinse in distilled water and then stain in P.T.A.H. overnight

5. Drain and rinse in 95% alcohol

6. Dehydrate, clear . Mount sections in DPX


Muscle striations, neuroglia fibres, fibrin, amoebae - Dark blue

Nuclei, cilia, red blood cells - Blue

Myelin - Lighter blue

Collagen, osteoid, cartilage, elastic fibres - Deep brownish red

Cytoplasm - Pale pinkish brown

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