DISTRIBUTION OF SUBSTANCES IN SPERMATOZOON. 239 



have a special affinity for lipoids, giving a salt by combination 

 with the lipoidic acid. After destroying the proteins by different 

 macerating reagents, one can isolate the lipoids and in this case 

 the staining with the mentioned substances is a specific stain. 



I shall not review this method in detail. Everyone who needs 

 to use it must read the original paper: Josef Schumacher: "Zur 

 Chemie der Zellfaerbung VIII. Mitteilung. Ueber die Nachweis 

 der Lipoide in Zelle und Gewebe," Chemie der Zelle und Gewebe, 

 Zeitschr. f. die Probleme der Gaerhung, Atmung u. Vitamin- 

 forschung, Bd. XII., Heft 5, 1926, S. 433. 



I have used his methods (see page 247) but most of the obser- 

 vations were made by simpler methods, using stains recommended 

 by him as specific stains, and in addition various intra vitam 

 staining methods. In the latter case the staining was done 

 under the cover slip as in the method of Koltzoff "Studien ueber 

 die Gestalt der Zelle," Arch. f. Zellforschung, Bd. II., S. i, 1909. 







II. RESULTS. 



i. THE USE OF STAINS WITH FRESH SPERMATOZOA. 



(a) Victoria Blue and Fuchsin. 



Victoria blue in i per cent, solution in distilled water: Stain 

 under the cover slip: one drop of sperm suspension (one drop 

 dry sperm to 10 cc. sea-water) is placed on the slide, and after 

 covering with a cover slip, with a fine pipette one adds at the 

 margin of the cover slip a drop of staining solution. The fluid 

 penetrates by capillarity, advancing more and more slowly. In 

 this way, one gets in the same preparation all degrees of staining, 

 from the strongest color to the complete lack of stain. At the 

 one end of the slide the spermatozoa are overstained, and on the 

 opposite side the spermatozoa are still active. 



The first impressive fact is a remarkable change of the shape 

 and size of the spermatozoa. The stain is in distilled water 

 and the spermatozoa are in sea-water: there is a great difference 

 in the osmotic pressure of the two fluids. Thus, a rapid penetra- 

 tion of the staining solution takes place into the spermatozoa, 

 which stain immediately. Shortly after, the heads of the 

 spermatozoa swell, become round and then burst; the following 



16 



