114 Manual of Veterinary 31lcrobioIoj>j. 



in absolute alcohol, the sections are cleared in oil of 

 cloves, passed into xylol, and finally mounted in bal- 

 sam. 



To obtain double staining by the violet method it 

 suffices to color the sections with picro-carmiue be- 

 fore placing them in the violet solution. 



Kiihne has modified this process so as to avoid the 

 employment of alcohol and oil of cloves. The sec- 

 tions, after dehydration in alcohol, are immersed for 

 ten minutes in the concentrated aqueous solution of 

 crystal violet, to which has been added hydrochloric 

 acid (one drop to fifty grams of the solution). They 

 are washed in distilled water, treated with Gram's 

 iodine solution, replaced in water, then passed rapidly 

 (a few seconds) through absolute alcohol and into 

 aniline oil in which they are decolorized. The ani- 

 hne is removed by xylol, and the sections mounted in 

 balsam. The employment of alcohol can be entirely 

 avoided by transferring the sections after the action 

 of the iodine on to the slide and there treating them, 

 after dehydration, with aniline, xylol, and balsam, in 

 succession. 



Method of Berlioz. — This gives a rapid double color- 

 ation. The sections are left for a quarter of an hour 

 in a mixture of equal parts of the two following 

 solutions : 



1. Distilled water, ... 84 cub. cent. 

 Aniline water, .... 6 " 

 Methyl violet 6 B, . . .2-5 grams. 

 Absolute alcohol, . . .10 cub. cent. 



2. Distilled water, ... 95 " 



90 per cent alcohol, . . .'5 " • 

 Coccbiine, 2-5 grams. 



