THE DISTRIBUTION OF SUBSTANCES IN THE SPER- 

 MATOZOON (ARBACIA AND NEREIS). 



STUDIED BY INTRA VITAM STAINS AND BY STAINS OF LIPOIDS 

 ACCORDING TO THE METHOD OF SCHUMACHER. 



DR. GREGOR T. POPA.i 



UNIVERSITY OF BUKAREST, ROUMANIA. 

 I. Introduction. 

 II. Results. 



1. The use of stains with fresh spermatozoa 239 



(a} Victoria blue and fuchsin 239 



(b) Janus green; trypan blue; neutral red 244 



2. Staining after fixation 246 



3. Schumacher's methods 247 



4. Osmic acid 249 



5. Sudan III 250 



6. Macrochemical test with Victoria blue 250 



7. Observations on living spermatozoa in sea-water 250 



8. The effects of egg-water on the spermatozoa 251 



III. Discussion 254 



I. INTRODUCTION. 



In a previous paper - I come to the conclusion that the sperma- 

 tozoa of mammals are surrounded by a layer of a lipoid substance 

 which is responsible for a reaction observed by Prof. Frank R. 

 Lillie, and called by me the lipo-gel reaction. 



With other methods and in different animal species, I have 

 been able to analyze much more completely the distribution of 

 this lipoid substance in and between spermatozoa. The following 

 description is a record of these new facts. 



In a recent paper Josef Schumacher has devised methods for 

 the identification of lipoids, lipoproteids and fats in the tissues 

 and cells by means of combining different strong reagents with 

 the action of stains. One conclusion of his work is that there is 

 a series of stains (above all Victoria blue and fuchsin) which 



1 From the Marine Biological Laboratory, Woods Hole, Mass. 



2 Gregor T. Popa, 1927, "A Lipo-gel Reaction Exerted by Follicular Fluid upon 

 Spermatozoa (Lillie's Reaction) and Its Significance, BIOL. BULL., Vol. 52, p. 223. 



238 



