Lillie's Short PAS Reaction
for 1-2 Glycols
The periodic acid Schiff reaction (PAS) is used to demonstrate the presence of 1-2-glycols, and is consequently an important method in the histochemistry of carbohydrates and the histological demonstration of many structures.
Lillie referred to this method as
The Periodic acid, Schiff Sulfite Leucofuchsin Reaction, short variant.
Periodic acid (potassium iodate, nitric acid specified).
Mayer's hemalum, or
Weigert's iron hematoxylin and saturated aqueous picric acid.
Sodium metabisulphite, 0.5% aqueous
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable.
Other fixatives are likely to be satisfactory, although glutaraldehyde should be avoided.
- Bring sections to water via xylene and ethanol.
- Oxidise in periodic acid for ten minutes.
- Wash in running water for five minutes .
- Place in Schiff's reagent for 10 minutes.
- Transfer to three successive baths of sodium metabisulphite
for 1, 2 and 2 minutes respectiively.
- Wash in running tap water for 5 minutes.
- Counterstain with one of the following:–
- Place in Mayer's hemalum for 2 minutes.
Wash in running tap water and blue.
- Place in Weigerts iron hematoxylin for 2-4 minutes.
decolorise with Pal's bleach diluted 1:5 with distilled water.
Wash in running tap water for 4 minutes.
- Place in Weigerts iron hematoxylin for 6 minutes.
Wash for 4 minutes in running tap water.
Place in saturated aqueous picric acid for 1 minute.
- Dehydrate with ethanol, clear with xylene and coverslip using a resinous medium.
- 1-2-glycols – red
- Nuclei – blue or black
- Cytoplasm – yellow or unstained
- The nuclear counterstain may obscure some positive staining. Keep the application time short.
- Weigert's solution should be allowed to ripen for an hour before use.
- Weigert's hematoxylin and Pal's bleach combination gives highly selective nuclear staining.
- Picric acid differentiates the nuclear stain as well as colouring the background.
- Glutaraldehyde fixed tissues will have a non-specific positive background staining.
This must be blocked before step 2.
Lillie, R.D., (1954)
Histopathologic technique and practical histochemistry Ed.2
Blakiston, New York, USA.
Last updated January 2019