Make your own Schiff's reagent - The world under the microscope

Schiff's reagent is used in, for example, the 'Feulgen reaction' or in the 'PAS' staining. Both are histochemical staining. In the Feulgen-reaction DNA is coloured and in the PAS (Periodic Acid-Schiff) colouring glycogen and other polysaccharides (carbohydrates) are used. The reagent can be bought ready to use but is quickly subject to decay and it is quite expensive.

For a large laboratory all less interesting, for the hobbyhistologist of course of great importance. Once the substances have been purchased, a desired quantity can be produced at any time.

The storage time can be greatly increased by storing the dye cool, dark and in a brown bottle. When the solution has become reddish it is no longer usable.

The recipe for 100ml reagent[1]:

Solution A: Dissolve 0.5gr pararosaline (1B-297[2]) in 15ml HCL 1N (a 1N solution contains 36.453gr hydrochloric acid per litre and therefore corresponds approximately to a value of 3.55%). With a 25% HCL solution, a dilution of 1ml HCL + 7ml AD gives a percentage of 3.57%. Attention: add acid to water and no water to acid!

Solution B: Dissolve 0.5gr potassium metabisulfite (K2S2O5) in 85ml AD.

Mix solutions A and B and leave to ripen in the dark for 24 hours (the reddish-brown colour slowly becomes more yellowish).

Pulverise 1 Tablet 'Norit' in a mortar and weigh 0,3gr.

On a magnetic stirrer, add the weighed activated carbon to the solution (do not heat the solution).

On a magnetic stirrer, add the weighed activated carbon to the solution (do not heat the solution).

Filter the solution.

Schiff's reagent is still rosa in colour but will be completely clear after a few hours.

Poured Schiff's reagent into a dark brown bottle.

The bottle should always have a sticker with the name of the product and a creation date.

The recipe as described in 'Romeis 18'[1].