SECTION TWO 



11. Expose the preparation to the air for a few seconds. 



12. Immerse for at least twenty minutes in solution F. 



13. Pass through the usual descending grades of alcohol into 

 distilled water. 



14. Proceed as steps 7-15, below. 



(b) Paraffin sections 



1. Fix sections to slides and remove paraffin wax with xylol 

 as usual. 



2. Wash in two changes of absolute alcohol. 



3. Coat with the celloidin solution (solution E). 



4. Expose the preparation to the air for a few seconds. 



5. Immerse in solution F for at least twenty minutes. 



6. Pass through the usual ascending grade of alcohol in 

 distilled water. 



7. Immerse in the acidified cresyl fast violet (solution D) for 

 three to ten minutes. 



Note: The solution can be used several times but it should be 

 renewed when signs of any loss of colour intensity are observed. 



8. Rinse in distilled water. 



9. Agitate the slides in 70% alcohol until the stain begins to 

 come out of the sections in clouds. 



10. Rinse quickly in 95% alcohol. 



1 1 . Immerse in chloroform for twenty minutes or more. 



12. Differentiate with 95% alcohol, controlling by microscopic 

 examination while the preparation is still wet, until the desired 

 coloration is attained. 



13. Dehydrate in absolute alcohol. 



14. Immerse in normal butyl alcohol (absolute) for one to five 

 minutes. 



14. Clear by immersion in at least two lots of xylol. 



15. Mount in D.P.X. or Clearmount. 



Notes: 



It is claimed that the technique offers at least two advantages 

 over other Nissl techniques : 



(a) Continuous exposure to light in the laboratory for almost 

 a year produced no noticeable fading or colour change. 



(b) The method greatly simplifies the staining of nerve cell 

 bodies in frozen sections. 



Reference: Fernstrom, R. C. (1958). 

 G 181 



