130 SOIL BACTERIOLOGY 



(f) Wash thoioughly with distilled water. 



C?) Dry and blot. 



Qi) 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. 

 (&) Air dry. 



(c) Fix by flaming. 



(d) Apply glacial acetic acid, drain immediately (So 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. 



