SECTION TWO 



8. Treat with 5% aqueous sodium thiosulphate for five minutes. 



9. Wash thoroughly in water for five minutes. 



10. Rinse in distilled water. 



11. Wash in 70% alcohol. 



12. Immerse in the aldehyde-fuchsin (solution E) for thirty 

 minutes to one hour, depending upon the age of the stain. 



13. Rinse in 95% alcohol. 



Note: It may be necessary to acidify the alcohol slightly to 

 remove background staining which will be present if the staining 

 solution has been used before it is fully ripe. 



14. Transfer to 1% periodic acid (solution F) and leave therein 

 for fifteen minutes. 



15. Immerse for two minutes in each of three changes of 

 distilled water. 



16. Immerse in the sensitized Schiff reagent (solution G) for 

 twenty minutes. 



17. Rinse in tap water. 



18. Immerse in acidified orange G (solution H) for ten minutes 

 to stain the acidiphils. 



19. Rinse quickly in water. 



20. Dehydrate through the usual graded alcohols. 



21. Mount in Michrome mountant, or proceed as follows: 



22. Clear in xylol. 



23. Mount in D.P.X., Clearmount, or Michrome mountant. 



Results: 



Thyrotrophs: deep purple. Gonadotrophs: red. Acidiphils: 

 orange. Nuclei: unstained. 



Notes: 



{a) It is stated that the absence of a nuclear stain greatly contri- 

 butes to the crispness of cytological detail. 



{b) The authors, who developed the procedure for routine 

 evaluation of rat and mouse pituitaries, state that it has also 

 proved useful for tissues in general, but for some tissues a counter- 

 stain other than orange G, which was found of particular value for 

 the pituitary, may be more useful. 



Reference: Elftman, H. (1959). 



121 



