SECTION SEVEN 



ANILINE BLUE - EOSIN B 



A simple and rapid technique for spermatozoa, which is 

 particularly suitable for dog and human semen 



Solutions required: 



Technique: 



1. Fresh semen is allowed to stand for about one half to one 

 hour until it liquifies. 



2. Prepare thin even smears of the liquified semen on scrupul- 

 ously clean and dry slides or coverslips. 



3. Fix for 3 minutes in a mixture consisting of equal volumes 

 of ether and absolute alcohol ; then allow to dry in the air. 



4. Flood smears with solution B and warm over a steam bath 

 or hot plate while the stain is allowed to act for 5-7 minutes at 

 40-60° C. 



5. Pour off excess stain. 



6. Wash the preparation thoroughly with distilled water. 



7. Drain and allow to dry thoroughly in air. 



8. Mount in a neutral synthetic resin such as D.P.X., Clear- 

 mount or Cristallite. 



Results: 



Sperm structure Stain reaction 



Galea capitis Pale bluish-grey; sharply outlined 



Cell membrane Bluish-grey; sharply outlined 



Nuclear membrane (shell) Bluish-grey ; sharply outlined 



Acrosome Slate blue 



Nucleas Pink 



Neck Sharply outlined (dark blue), inside 



colourless 



347 



