SECTION TWO 



6. Cedarwood oil, one to sixteen hours. 



7. Xylol, for fifteen minutes. 



8. Infiltrate in paraffin wax 56-58° C. (four changes before 

 finally embedding). 



9. Fix sections, 4jLt in thickness, to slides and remove paraffin 

 wax by immersing in two changes of xylol for three minutes each. 



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

 absolute alcohol. 



11. In 95% alcohol for three minutes. 



12. Distilled water for three minutes. 



13. Lugol's Iodine for three minutes. 



14. Sodium thiosulphate solution for three minutes when the 

 sections should have been restored to their natural colour. 



15. Stain in Delafield Haematoxylin for thirty seconds. 



16. Wash in tap water for three minutes. 



17. Immerse in distilled water for three minutes. 



18. Immerse in 80% alcohol for three minutes. 



19. Aniline alcohol (Solution E) for fifteen minutes. 



20. Stain in Azocarmine for forty-five minutes. 



21. Rinse in distilled water. 



22. Differentiate in aniline alcohol for two to three minutes. 



23. Wash in acid alcohol (solution G) for thirty to sixty seconds. 



24. Immerse in phosphotungstic acid solution for one hour. 



25. Dehydrate by passing through 70%, 95% and absolute 

 alcohols (two minutes in each). 



26. Counterstain in acid green - orange G solution for five 

 minutes. 



27. Clear in xylol for one minute. 



28. Immerse for half an hour in each of two changes of xylol, 

 to remove completely all traces of clove oil which would otherwise 

 cause further decolorisation. 



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



Results: 



Alpha granules, purplish red. Beta cell granules, light green. 

 Nuclear membranes are sharply defined and mitochondria are 



69 



