444 WILLIAM M. GOLDSMITH 
Cicindela repanda, C. purpurea, C. ancocisconensis, and C. 
vulgaris emerge in late summer or early fall, hibernate, and reap- 
pear in the spring, they are unquestionably classed under type 
‘a’ and thus double-brooded. Specimens dissected from fall 
collections of these species proved very immature, but were very 
favorable for a study of the early spermatogonia and oogonia. 
On the other hand, the spring collections from these double- 
brooded forms were found to be of great value for a study of the 
later stages of the germ cells. 
Since Cicindela punctulata and C. sexguttata appear in Indi- 
ana in late spring or early summer and die about eight weeks 
later, they are spoken of as single-brooded. Shelford’s observa- 
tions on the larvae and pupae of the latter form indicate that 
this species is in reality double-brooded. However, the young 
adults of the northern range do not dig their way out in the fall, 
but remain in the pupal burrows until spring. Since the single- 
brooded forms mature at once after appearance, there is a range 
of only about three or four weeks in each year in which favor- 
able cytological material can be collected. 
The technical side of the study of Coleopteran cytology is 
very difficult and disappointing in its results. All fixations in 
common use were tried under various conditions, but none 
proved entirely satisfactory. Several hundred specimens of 
Cicindelidae were dissected, resulting in but a few good fixations 
from each species. All of the best preparations were fixed in 
Flemming’s fluid (strong). This method gave best results when 
the warm solution was dropped into the body cavity of the live 
specimen before dissecting out the gonads. After the fluid had 
had time to penetrate slightly all parts of the body, the gonads 
were removed and placed in cold Flemming for two hours. Even 
under these conditions good fixations were exceptional, and no 
explanation could be given for a good preparation when obtained. 
One of the most perfect fixations (C. sexguttata) was one of 
seventeen specimens collected, dissected, and treated under the 
same conditions. The other sixteen were absolutely worthless. 
Iron-haematoxylin, with orange G as a counterstain, when 
needed, was used most extensively. 
