Place the phenol in an Erlenmeyer flask and melt it under hot water through the glass.
Add the chromotrope 2R and mix well into a sludge. Add the water and mix well. Filter before use.
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable.
Other fixatives are likely to be satisfactory.
Bring sections to water via xylene and ethanol.
Stain nuclei with Mayer's hemalum and blue.
Place in the staining solution for 30 minutes.
Wash well with tap water.
Dehydrate with ethanol.
Clear with xylene and mount with a resinous medium.
Nuclei – blue
Eosinophil granules – bright red
Erythrocytes – may be lightly stained
Paneth cell granules – may be brownish
Enterocromaffin granules – may be brownish
The basis of this method is difficult to rationalise. Phenol is acidic and thus lowers the pH. This is often used as the basis for explaining the method, but usually an acid dye stains all basic components of the tissue (muscle, cytoplasm, collagen) intensely when applied at an acid pH. With this method eosinophils are intensely stained, but other components that would usually stain with an acidified acid dye are not. Phenol can have an intensifying effect, as is clear from its inclusion in carbol fuchsin, when it intensifies staining with basic fuchsin, but the mechanism has not been satisfactorily explained.
Reference Histological demonstration techniques, (1974)) Cook, H C.
Butterworths, London, England.