CYTOPLASMIC INCLUSIONS OP THE GERM-CELLS. 563 
blasted into fragments by many so-called “ admirable fixa- 
tives.” 
In this work on the snail I used the following fixatives : 
(1) My modification of Flemming’s strong formula, in 
whicn no acetic was used. 
(2) The same fluid diluted one-sixth. 
(3) Champy’s fluid. 
(4) Flemming’s strong solution, in which the acetic acid 
was re-placed by nitric acid. 
All the better-known fixatives were also tried, such as 
Flemming, Perenyi, corrosive sublimate, Carnoy, etc. 
For staining I used the usual stains, such as Ehrlich’s 
heematoxylin and eosin, Mayer’s acid heemalum, and iron 
alum hsematoxylin with orange G., Bensely’s acid fuchsin- 
methyl green, and alizarin-toluidin blue, etc. 
Taking the reddish or orange alizarin or acid fuchsin as a 
basis, the slides were often counterstained in toluidin blue, 
crystal violet, methyl green, thionin, etc., and the same was 
done taking iron heematoxylin as the first stain. Some slides 
were mounted unstained in euparal for studying yolk. 
Smears were tried, but failed to give very helpful results 
except for studying later stages and ripe spermatozoa. 
Of the large number of fixing and staining methods used, I 
found that for the snail the following was the best : The 
animals are anaesthetised in chloroform vapour, and as soon 
as possible the last upper whorls of the shells are carefully 
broken with strong forceps or by a blow with some hard 
instrument. The shell is dipped in water, and the remaining* 
pieces around the broken area are removed with forceps. 
The ovotestis is cut out and laid on the table and superfluous 
digestive gland is cut away, exposing the milk-coloured 
ovotestis on every side. This piece is cut in half longi- 
tudinally, and immediately thrown into a capsule of Flemming 
without acetic acid, diluted one-sixth with distilled water. 
Here the fragments are left overnight. They are washed for 
at least two hours in running water next morning, and then 
brought up from 50 per cent, alcohol and the other grades to 
