156 PATHOGENIC BACTERIA. 



Method of Staining Spores. — It has already been 

 remarked that the peculiar quality of the spore-capsules 

 protects them from the influence of stains and disinfect- 

 ants to a certain extent. On this account they are much 

 more difficult to color than the adult bacteria. Several 

 methods are recommended, the one generally employed 

 being as follows : Spread the thinnest possible layer of 

 material upon a cover-glass, dry, and fix. Have ready 

 a watch-crystalful of Ehrlich's solution, preferably made 

 of fuchsin, and drop the cover-glass, prepared side down, 

 upon the surface, where it should float. Heat the stain 

 until it begins to steam,, and allow the specimen to 

 remain in the hot stain for five to fifteen minutes. The 

 cover is now transferred to a 3 per cent, solution of hydro- 

 chloric acid in absolute alcohol for about one minute. 

 Abbott recommends that the cover-glass be submerged, 

 prepared side up, in a dish of this solution and gently 

 agitated for exactly one minute, then removed, washed 

 in water, and counter-stained with an aqueous solution 

 of methyl or methylene blue. 



In such a specimen the spores should appear red, the 

 bacilli blue. 



I have not generally found that spores color so easily, 

 and for many species the best method seems to be to 

 place the prepared cover-glass in a test-tube half full of 

 carbol-fuchsin : 



Fuchsin, 1 ; 



Alcohol, 10 ; 



5 per cent, aqueous solution of phenol crystals, 100, 



and boil it for at least fifteen minutes, after which it is 

 decolorized, either with 3 per cent, hydrochloric or 2-5 

 per cent, acetic acid, washed in water, and counter- 

 stained blue. 



Muir and Ritchie 1 recommend that cover-films be pre- 

 pared and stained as for tubercle bacilli, then decolorized 

 with a 1 per cent, sulphuric acid solution in water or 



1 Manual of Bacteriology, London, 1897. 



