ANJESKY'S METHOD OF STAINING SPORES 91 



carbol-f uchsin ; methylen blue, aqueous^alcoholic ; hydro- 

 chloric acid, 0.5%; sulphuric acid, 5%; Canada balsam; 

 microscope. 



Culture. Culture of an organism just beginning to 

 show spore formation. 



Method. 1. Prepare a cover-glass film of the spore- 

 containing organism and allow it to dry. 



2. While it is drying, warm some 0.5% HC1 in the small 

 evaporating dish over a Bunsen burner until it steams 

 well and bubbles begin to form. 



3. When the solution is hot and the smear dry, drop 

 the cover-glass upon the fluid and allow it to act upon the 

 unfixed smear for three to four minutes. 



4. Wash and dry the cover-glass. 



5. Fix in the flame for the first time. 



6. Stain with carbol-fuchsin by flooding the cover- 

 glass with the stain, warming twice until fumes arise. 



7. Allow to cool, and wash in water. 



8. Decolorize with 5% H 2 SO 4 . Spores are treated with 

 a mild decolorizing agent, as they are much less resistant 

 to acid than are acid-fast bacteria. (See p. 93, step 7.) 



9. Wash in water. 



10. Counterstain for one to two minutes with methylen 

 blue. 



11. Wash, dry and examine the specimen in water. 

 If satisfactory, dry it and mount in balsam. 



The whole procedure should not take longer than eight 

 to ten minutes. 



REFERENCE 



MCFARLAND: Textbook of Pathogenic Bacteriology, p. 188. 



