Gram-Weigert and Verhoeff Carbol Fuchsin

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From:Gayle Callis <>
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Whew, this was more fun than dumpster diving!!  Glad I horde old books!

Both of these methods are in Lillies book, the Gram Weigert is a variation
of the original, and is listed as the Weigert Fibrin Stain, same for the
method in Gurr, older and the one I have here is the Gurr, and probably the
original, with all the aniline oil, although results should be the same.
Lillies book gave an extensive explanation of the method, plus history and
notes on the stain. 

Gram Weigert Method

Delafield Hematoxylin

Aniline gentian violet

gentian violet (crystal?) 5 g
aniline oil   2 ml
distilled water 88 ml
alcohol, 95%    10 ml

Lugols iodine

aniline oil  20 ml
xylene       10 ml

erythrosin 5% in absolute ethanol

fix with NBF
bring to water, stain in Delafields for 5- 20 min
quick rinse in acid alcohol
rinse, tap water 2-5 min
aniline gentian violet for 2-5 min
pour off excess stain, blot slide WITHOUT WASHING
flood with Lugols, 2-5 min
pour off excess and blot without washing
decolorize for a few seconds with aniline oil/xylene
flood with erythrosin solution 30 sec to 1 min
wash this off with aniline oil/xylene, then rinse with xylene, drain and
blot dry carefully, mount

From Gurr, Staining Practical and theoretical, 1962 William and Wilkins, 

Ref:  Weigert, C (1887)Fortschr. Med, 5:228-232.
      Mallory FB (1938) Pathological technique, WB Saunders London)p 272 

Verhoeffs Carbol Fuchsin is in Lillies book, 4th Edition

26.8 g phenol or 25 ml melted crystals, in 50 ml absolute ethanol.  
add 2 g basic fuchsin, heat at 37C with occasional shaking for 18-24 hr,
Filter and store

Working solution:  1 ml stock in 6 ml water
Lillie, p 736, Chapter on Smear, bacteria protozoa and other parasites. 

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

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