No. 1. — Spermatogenesis of Caloptenus femur-rubrum and Cicada 
tibicen! By E. V. WILCOX. 
Tun following observations were made on the testes of Cicada and 
Caloptenus, Only three male Cicadw were at my disposal, but of 
Caloptenus I examined more than twenty individuals, The Cicade 
were killed immediately after leaving the pupal case, and had been pre- 
served a number of years. The Calopteni were taken in August and 
September, 1893. 
'l'he testes of Cicada were killed in Müller's fluid ; those of Caloptenus 
either in hot water, in hot corrosive sublimate, in cold corrosive subli- 
mate, or in chrom-osmic-acetic mixture. Some of the testicular folli- 
cles of Cieada were stained in Grenacher's aleoholie borax carmine, 
others according to Bizzozero’s modification of Gram's method. The 
follicles were stained in toto in safranin (50% alcohol) 24 hours, sec- 
tioned, stained 3 minutes in gentian-violet, washed 5 minutes in a solu- 
tion of potassic iodide, then treated alternately with alcohol and chromic 
acid (0.1%). But better results were obtained by double staining with 
safranin and victoria-green. Crystals of the latter were dissolved in 
absolute alcohol, or in clove oil. The sections were first stained in 
safranin (10—15 minutes), tho excess of stain being quickly washed off 
in 90% alcohol, and then in a very strong solution of victoria-green in 
absolute alcohol for 1 to 2 minutes. Staining and dehydrating were 
thus accomplished at the same time. Excess of green was washed out 
with absolute alcohol. Sections were cleared in clove oil When a 
clove-oil solution of the green was used, the sections were dehydrated 
before staining in the green. The method with the absolute-alcohol 
solution gave the better results, and was more easily managed. 
The Caloptenus material was all stained on the slide. The methods 
used were either safranin and victoria-green, as just described, Henne- 
guy’s (91) potassic permanganate and safranin, or Heidenhain’s (92) 
iron-heematoxylin. "The method with safranin and victoria-green gave 
good results,  COytoplasm and achromatic nuclear parts were stained 
green, the chromosomes, nucleolus, and centrosomes red. If the green 
1 Contributions from the Zoölogical Laboratory of the Museum of Comparative 
Zoology at Harvard College, under the direction of E. L. Mark, No. XLVII. 
VOL, XXVII. — NO. 1. 
