804 Stoward,—A myloclastic Secretory Capacities of the 
Immediately after removal of the objects from the copper sulphate 
solution, they were divested of their paleae, and either washed as thoroughly 
as possible with sterilized tap-water only or washed and re-steeped in tap- 
water for 24 to 48 hours. 
When absolute alcohol served as the antiseptic steeping reagent this 
re-steeping in water was invariably resorted to, and subsequent experience 
demonstrated the value of employing two to three changes of water or of 
placing the seeds under aeration conditions for 24 hours after removal 
from the water-steep. 
For special purposes endosperms and inner endosperms prepared from 
air-dried seeds were successively steeped in absolute alcohol and water. In 
every instance the volume of steeping liquid used was just sufficient to 
cover the material. Small well-stoppered weighing bottles were employed, 
and each phase of the operation was carried out under strictly aseptic 
conditions. 
The efficiency of the reagents employed and the method of manipulation 
followed was amply foreshadowed by the results furnished by several series 
of preliminary experiments. 
Seeds treated as described were placed in tubes containing various 
nutrient media, and were then incubated at 30° C. for 7 to 14 days. In the 
majority of instances the contents of the tubes remained absolutely sterile :— 
in a small percentage only, moulds, but not bacterial forms developed, thus 
demonstrating the fact that the spores of the former organisms alone survived 
the toxic action of the reagents employed. 
Preparation of sterilized objects for cultural purposes and investigation 
of their relative amylolytic secretory or generative capacities. The dissection 
of the steeped sterilized material was carried out in a Hansen inoculation 
chamber under the following aseptic conditions :— 
A flat glass plate was placed on the floor of the chamber (the interior of 
which, just prior to use, was thoroughly swabbed with 1 % HgCl 2 solution) 
and covered with an inverted glass funnel. The depaled, steeped, thoroughly 
washed grains were then introduced en masse under the funnel. 
Each grain was then successively withdrawn, dissected as required, and 
returned under the funnel until a sufficient number of objects for a culture 
experiment were prepared. 
Dissections of grains into embryos and endosperms were accomplished 
by cutting through the integuments in the neighbourhood of the scutellar 
margin and gently lifting out the embryo with a scalpel: the further dis¬ 
section of the endosperm into inner endosperm and aleurone layer was carried 
out by carefully cutting off the latter with a sharp razor in such a manner 
as to include the merest traces only of the subjacent amyliferous tissue. 1 
1 This was subsequently controlled by filing off the aleurone layer of a number of air-dried 
grains and comparing the dry weights of this material after desiccation for 24 hours at 30° C., with 
