Lillie's
Schiff Reagent

Solution

Ingredient Amount
Pararosanilin 1 g
Distilled water 100 mL
Potassium metabisulphite 2 g
Hydrochloric acid, 1N 20 mL
Activated charcoal (powder) 0.3 g

 

Preparation
Bring the water to boiling.
Remove from the heat and add the dye (be careful of foaming).
Cool to 60° C and filter.
Add the metabisulphite and the hydrochloric acid and mix well.
Leave overnight or longer.
Add the activated charcoal, shake well for about a minute and filter.
Store at 0-5°C in a tightly stoppered bottle.

 

Notes

  1. Lillie specified sodium or potassium bisulphite or metabisulphite. According to the Merck Index, commercial sodium bisulphite is made predominantly of sodium metabisulphite, and for most purposes they are interchangeable.
  2. Lillie specified the dye as pararosanilin, rosanilin or new fuchsin. Pararosanilin is now known to produce the most consistent Schiff's reagent, the other dyes being less satisfactory.
  3. The solution should be colourless or very pale amber. Brown solutions will often stain poorly. Usually these have been made from a batch of basic fuchsin containing too little pararosanilin.

 

Reference
Lillie, R.D., (1954)
Histopathologic technique and practical histochemistry Ed.2
Blakiston, New York, USA.

 


 

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