XI. STAINING THE GLANDERS BACILLUS. 



(A) IN COVER-GLASS PREPAEATIONS. 

 a. LOEFFLER'S OLD METHOD. 



1. Flood the cover-glass for three minutes with Loeff- 

 ler's methylene-blue solution (See XIV, foot-note 2). 



2. Heat in the flame until the fluid steams (do not 

 boil). 



3. Wash in water. 



4 Wash in acetic-acid water (1 :100) from five to ten 

 seconds. 



5. Wash in water. 



6. Mount, etc. 



b. LOEFFLER'S NEW METHOD. 



1. Flood the glass with alkaline aniline-water-gen- 



tian-violet solution,^ five minutes. 



2. Wash in acetic-acid water (1 :100) rendered a Rhine- 

 wine-yellow color by the addition of a few drops of 

 tropaeolin (oo) solution. 



3. Wash in water, mount, etc. 



c. A SIMPLE METHOD. 



1. Flood the cover-glass with 2 per cent, gentian-violet 

 solution (See under I, foot-note 2 a). 



2. Heat in the flame untU the fluid steams well (do 

 not boil), and allow it to stand from one-quarter to one- 

 half minute. 



3. Wash in water, mount, etc. 



(B) IN SECTIONS, 

 a. LOEFFLER'S METHOD. 



1. Ntain for a few minutes in LoefHer's methylene- 

 blue solution (See XIV, foot-note 2). 



2. Five seconds in the following solution : Aq. dest., 

 lOcc ; concentrated H2SO4 , 2 drops ; 5 per cent, oxalic acid, 

 1 drop. 1 ! 



3. Decolorize and dehydrate in absolute alcohol; oil; 

 balsam. I 



lAlkaline aniline-'water-sentian-Tiolet solution (See III, foot-note 

 2, a, b) mixed with an eqnal amount of a potash solution (1:10,000), 

 or an equal amount of Liq. Ammon, caustic, half per cent. 



35 



