DIRECTIONS FOR THE USE OF STAINS I2Q 



7. The mount should be labeled as follows: Name of 

 organism, stain, and date : 



& o 



1. Gram's Stain: 



(a) Spread an even, thin film. 



(b) Fix in the flame. 



(c) Apply aniline oil .gentian- violet for two to five min- 



utes. 



(d) Wash in water. 



(e) Apply Gram's iodin for one minute, or until the 



preparation has a coffee color. 

 (/") Wash in water. 

 (g) Decolorize with 95 per cent, alcohol until no more 



violet color streams out. 

 (ti) Wash in water. 



2. Endospore Stain (Double): 



(a) Spread thin film. 



(b) Fix by passing through flame three times. 



(c) Stain with hot carbol-fuchsin five minutes (do not 



boil). 



(d) Clean under side of slide with 2\ per cent, acetic acid. 



(e) Decolorize the smear with i\ per cent, acetic acid 



until the pink color is nearly removed from film. 



9 



