216 TECHNIC, 



be used in simple staining and in connection with 

 other stains. 



II. STAINING METHODS. 



(1) SIMPLE STAINING. 



This consists in using a single stain. 



(2) DOUBLE STAINING. 



This consists in using two stains, one to stain spores, 

 protoplasm, etc., and the other as a ground stain. The 

 following methods will illustrate double staining: 



STAINING OF SPORES. 



(a) Make a cover-glass spread, dry and pass three 

 times through the flame. 



(b) Add aniline- water gentian -violet. 



(c) Heat until the preparation begins to boil; re- 

 move for a minute. Repeat this process six times. 



(d) Wash in three per cent hydrochloric acid-alcohol 

 one minute. 



(e) Wash in water. 



