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FIXATION:        10% buffered neutral formalin.

TECHNIQUE:        Paraffin sections cut at 5 μm.

SOLUTIONS:        

Weigert’s Iron Hematoxylin Solution

(see Masson’s Trichrome Stain)

Van Gieson’s Solution

Acid fuchsin, C.I. 42685, 1% aqueous ------------------------------------------                4.0 ml

Picric acid, saturated aqueous --------------------------------------------------                40.0 ml

1. Deparaffinize and hydrate to distilled water.
2. Weigert’s iron hematoxylin solution for 5 minutes.
3. Wash briefly in running tap water and rinse in two changes of distilled water.
4. Van Gieson’s solution for three minutes.
5. Dehydrate in three changes of 95% alcohol (Coplin jars) and complete dehydration in three changes of absolute alcohol.
6. Clear in three or four changes of xylene.
7. Mount with synthetic resin.

Collagen -------------------------------------------------------------        red

Muscle ---------------------------------------------------------------        yellow

Cornified epithelium -------------------------------------------------        yellow

Nuclei ----------------------------------------------------------------        blue-black

The main advantage of the Van Gieson stain is that it provides for a sharp contrast between collagen (red) and everything else (yellow).  It is also quite useful as a counterstain in various other procedures, i.e., Verhoeff’s and Iron Gallein elastic methods, especially when these are used in tissue such as cirrhotic liver.

Mallory, F.B.:  Pathological Technique, New York, Hafner, 1968, pp. 92-93, 152-153.