SECTION TWO 



D. Erythrosin . . . . . . i gm. 



Tap water . . . . . . loo ml. 



Note: If this solution is to be kept in stock for 

 more than a few days, add a few drops of chloroform 

 to prevent decomposition by air-borne micro- 

 organisms. 



Technique: 



1. Fix adult pituitaries in Bouin's fluid or in io% formalin 

 and embed in paraffin wax as usual. 



2. Cut sections at 5/x. and attach them to slides. 



3. Dewax with xylol. 



4. Rinse with absolute alcohol. 



5. Wash with two changes of 95% alcohol. 



6. Immerse sections in the luxol fast blue (solution A) in an 

 incubator at 55° C. for two to four hours. 



7. Wash off excess stain with 95% alcohol. 



8. Rinse with distilled water. 



9. Begin the differentiation by immersing the preparation in 

 0-05% lithium carbonate (solution C) for three to five seconds. 



Note: Considerable care has to be exercised to obtain the 

 correct degree of differentiation as this is a very delicate process. 



10. Continue the differentiation in four changes of 70% alcohol. 

 Note: The final lot of alcohol should be free of stain. 



11. Wash with distilled water. 



12. Check the degree of differentiation by microscopic exam- 

 ination, and if further differentiation is necessary, repeat steps 10 

 and II quickly. 



13. Wash preparations in tap water. 



14. Counterstain in the erythrosin (solution D) for five to ten 

 seconds. 



15. Wash in tap water. 



16. Wash in 90% alcohol. 



17. Dehydrate in absolute alcohol. 



18. Clear in xylol. 



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



Results: 



Alpha cells: blue. Colloid: pale blue. 



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