STAINING OF BACTERIA IN TISSUES. J 53 



of either eosin, Bismarck-brown, or safranin for one 

 minute, then washed out in alcohol, finally in absolute 

 alcohol, and then into xylol, from which they may be 

 mounted in the manner given. 



Especially brilliant results are obtained when tissues 

 containing anthrax bacilli are stained by this process ; 

 the bacilli will be of a deep blue color, while the sur- 

 rounding tissues will be of the color used as contrast. 



Kuhne's carbolic methylene-b/ue melhod. Stain the 

 sections in the following solution for from one-half to 

 one hour : 



Hethylene-blue, In substance .... 1.6 erammes. 

 Absolute alcohol 10 c.c. 



Rub up thoroughly in a mortar, and when the blue 

 is completely dissolved, add gradually 100 c.c. of a 5 

 per cent, solution of carbolic acid. (The solution de- 

 composes after a short time ; it should be made fresh 

 when needed.) From this the sections are washed out 

 in water, then in 1.5 to 2 per cent, hydrochloric acid in 

 water, from this into a solution of lithium carbonate of 

 the strength of six to eight drops of a concentrated 

 watery solution of the salt to ten drops of water, and 

 from this again thoroughly washed in water ; then into 

 absolute alcohol containing enough methylene-blue in 

 substance to give it a tolerably dense color, then for a 

 few minutes into aniline oil to which a little methylene- 

 blue in substance has been added; then completely 

 rinse out in pure aniline oil, from this into thymol or 

 oil of turpentine for two minutes, and then into xylol, 

 from which they are mounted in xylol-balsam. The 

 advantages of this method are that it is generally ap- 

 plicable, and by its use the bacteria are not robbed of 

 their color, whereas the tissues are sufficiently decolor- 



