472 HOPE HIBBARD 



and Flemming without acetic acid. In the last-named fluid 

 one series was fixed for one day and another for seven days. 

 The fixation in Perenja and Cajal was very poor, and therefore 

 the material was discarded. All the material was imbedded at 

 Woods Hole in soft paraff.n, then taken to Bryn Mawr, reim- 

 bedded, sectioned, and stained. The sections were for the most 

 part 4ju in thickness. 



The stains employed were Heidenhain's iron hematoxyhn, 

 Auerbach's acid fuchsin-methyl green, hthium carmine and Lyons 

 blue, basic fuchsin and methylene blue, Benda's ahzarin and 

 crystal violet, safranin, safranin and gentian violet and orange 

 G, and for special tests, sudan III and Ziehl's carbol-fuchsin. 

 Samples of material fixed in solutions containing osmic acid were 

 also mounted unstained. Of these stains, the iron hematoxylin 

 and the Benda stain proved the most satisfactory and were the 

 most widely employed. In making up the alizarin the direc- 

 tions given in Guyer's Animal Micrology were followed rather 

 than those given by Benda himself or by Cowdry in describing 

 Benda's method. Guyer gives the following formula for Benda's 

 solution of sulphaUzarinate of soda: 1 part of saturated aqueous 

 solution of stain to 80-100 parts of water. Benda's own direc- 

 tions are to add 1 part of a saturated alcoholic solution of the 

 stain to 80-100 parts of water. Both methods were tried but 

 Grlibler's sulphaUzarinate of soda was found to be practically 

 insoluble in alcohol. The stain made from the saturated aqueous 

 solution of the dye gave excellent results. 



OBSERVATIONS 



As has been mentioned above, there were no visible differences 

 between the self-fertilized and the cross-fertihzed eggs. Any 

 given method of fixation followed by the same stains gave identi- 

 cal results in the two cases. In order to compare them the better, 

 sections of self-fertihzed eggs and sections of cross-fertihzed eggs 

 were mounted side by side on the same sHde. This insured ex- 

 actly the same degree of staining. 



A number of structures were found in the cytoplasm following 

 different methods of fixation and staining. All the bodies 



