Hi; University of California Publications. [zoology. 



Technique. — For this investigation, material has been col- 

 lected each mouth, and twice in the month during the spring, when 

 the tides were favorable. Although specimens have been secured 

 from various localities, they have been regularly obtained from 

 a locality known as Lands End, near the entrance of the Golden 

 Gate, California. The results most relied upon have been 

 obtained then, from material killed and fixed under the most 

 favorable circumstances, i.e., very soon after collection. The 

 relatively thick calcareous ectocyst of Grisia makes it difficult 

 to fix the tissues rapidly enough to prevent their shrinkage and 

 consequent distortion. The most successful results were obtained 

 by the use of a solution of hot corrosive sublimate. In most 

 cases a solution of this with glacial acetic was used, in other 

 cases, the hot corrosive sublimate alone. The specimens were 

 allowed to remain in the fixing fluid only long enough to become 

 penetrated, when they were washed in 50 % alcohol containing 

 iodine. After this, they were carried through the various 

 grades of alcohol and finally preserved in 85% alcohol. The 

 process of killing aud fixing did not include decalcification. 

 Such portions only as were required for mounting, were afterward 

 completely decalcified. In the process of decalcification, much 

 trouble is frequently experienced by the formation of bubbles of 

 gas. It was found easy to avoid these, however, and the conse- 

 quent tearing of the tissues, by decalcifying small pieces in a 

 high grade of alcohol made weakly acid. The stains used were 

 Delafield's and Ehrlich's hematoxylin with eosin; Benda's iron 

 hii'inatoxylin alone, and with eosin and fuchsin; and Auerbach's 

 mixture of methyl green and fuchsin. Many other stains were 

 experimented with, but these gave the most satisfactory results. 



Four species of Grisia are more or less abundant in this 

 region, viz., Grisia eburnea, Crisia geniculata, Grisia cornuta, 

 and a new species, Crisia occidentalis. A full description of this 

 last species will follow in a later paper. Special reference is 

 made in this paper to Grisia eburnea, although all the species have 

 been studied more or less in regard to their method of reproduc- 

 tion. Grisia eburnea is certainly dioecious, the two kinds of 

 genital products never being found in the same colony. This is 

 thought to be true also of Crisia nrrith nfalis, although the evi- 



