and Laboratory Methods. 



1779 



Fig. 8. — Two living spermato- 

 zoids in the mother cell, the 

 spiral bands being turned to- 

 ward the observer ; about 

 x200. 



unmistakable tail-like appendage. Is Hirase's spermatozoid an abnormal form 



or an injured one ? The latter alternative seems to be more probable. Mr. 



Fujii observed a spermatozoid form a tail-like ap- 

 pendage by pressure or some other cause, and also 



such a deformed spermatozoid resumed a normal 



form by absorbing the tail. I have also found a 



normal spermatozoid protruding a tail-like process 



before its death. 



The spiral band on the spermatozoid seems to 



be of two and a half turns instead of being three, 



as described by Mr. Hirase and followed by Mr. 



Bessey (Fig. 8). In this point Mr. Fujii also 



agrees with me. 



For examining the sperniatozoids, first cut the 



ovule into two parts, and from the upper half take 



off the fleshy outer and hard inner integuments. 



Then a thin paper-like nucellar cap is exposed. 



On the somewhat pointed apex of it make a shallow^ 



cut about 3-4 mm. square with a sharp knife or razor, and peel off the part with 



a pair of forceps. Now in the square area 

 of pale greenish endosperm which is thus 

 exposed, a central process and two (some- 

 times three or four) small concavities will be 

 seen. These holes are the entrance into 

 the archegonia imbedded in the tissue of 

 the endosperm. On the underside of the 

 peeled piece of nucellus, one to several 

 shining bodies may be seen. These bodies, 

 very minute and resembling miniature dew 

 drops, are the pollen-tubes. 



Now put the square piece of the nucel- 

 lus already peeled off, with the outer surface 

 down, on a slide, and add a drop of cane 

 sugar solution (about 6-8 per cent.)^ before 

 placing on a cover-glass. Examined under 

 the microscope, a large cell with two sper- 

 matozoids inside may be seen at the free 

 end of the pollen-tube (Figs. 7, 8). Some- 

 times instead of the spermatozoids we can 

 see only a single oblong-elliptical or fusi- 

 form body, the nucleus, in the center of the 



Fig. 9.-Spermatozoids swimming free ^g^ .pig. 3). This is the Stage before the 

 m pollen-tube (somewhat diagram- . 



atic); about X 200. division of the cell to form the sperma- 



I. Dr. Webber used in his studies on Zamia 10 per cent, solution with a good result. I 

 found that 10 per cent, seems to be slightly strong for Gin /ego, 6-8 per cent, being seemingly a 

 right concentration. 



