130 SOIL BACTERIOLOGY 



(/) Wash thoroughly with distilled water. 



(g) Dry and blot. 



(h) Counterstain with Loffler's methylene-blue for ten 



seconds, 

 (i) Wash in water, mount, and examine. 



3. Capsule Stain: 



(a) Spread film without the use of water. 



(b) Air dry. 



(c) Fix by flaming. 



(d) Apply glacial acetic acid, drain immediately (clo not 



wash in water). 



(e) Wash off acid with carbol-fuchsin three times, as 



rapidly as possible. 

 (/") Wash in i per cent, salt solution. 

 (g) Mount in the salt solution. 



4. Flagella Stain (Preparation of Culture): 



(a) Transfer cultures twice each day for five days; in 



the morning into bouillon, and in the evening on 

 to fresh slopes of bouillon agar. 



(b) Carefully inoculate a tube of sterile city water. 



(Slant the tube and make the inoculation at the 

 base. Then gently raise tube to upright position.) 

 Incubate for one hour at 37 C. 



(c) Make smears from the top of the water culture, 



using the greatest care in each step to prevent the 

 breaking of the flagella. 



(d) Air dry. 



(e) Flame once only. 



