23 
Note III .—The longer a film is washed the more 
blue comes out of the red cells which eventually 
become pink, but at the same time the red chromatin 
stain of the nuclei of parasites, etc., is also partly 
dissolved. Blue may be extracted more readily by 
using dilute spirit. A film that has been washed too 
much or insufficiently stained can easily be restained 
(first dissolving off cedar wood oil with xylol). 
2. Leishmari* s Stain .—Is the precipitate resulting 
from the action of a watery solution of alkalised 
methylene blue on a watery solution of eosin. The 
precipitate is filtered off, washed and dried. It is then 
dissolved in pure methyl alcohol. 
(Leishman’s stain in 4 soloids ’ 0*015 grammes, 
methyl alcohol 10 c.c.; or the solution may be 
bought ready made, but its keeping properties are 
doubtful.) 
Jo Stain. — (a) Without fixing, with a pipette, 
cover the film with the solution for one minute. 
(b) Add about twice the quantity of water and 
mix carefully on the slide (with the pipette). Allow 
to stain five to ten minutes or for hours. 
( c ) Wash in water. 
3. Giemsa Stain .—From the alkalised solution of 
methylene blue, Giemsa prepares in a pure condition 
some of the derivatives, e.g., methylene azure (Azur I). 
An impure form of this, consisting of methylene azure 
and methylene blue in equal parts, he calls Azur II, 
and in fact it is this body that he uses in his stain, as the 
presence of methylene blue is necessary in order to 
stain the cell body. Further, he mixes with this an 
impure substance, Azur II-Eosin—which is the 
precipitate got by the action of eosin on alkalised 
methylene blue and which, therefore, consists of 
Azur I-Eosin, methylene blue-Eosin, with some methy- 
