SECTION TWO 



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 

 orange red. Erythrocytes : brilliant orange. Golgi apparatus shows 

 as negative image in both alpha and beta cells. Chromophobes show 

 little or no cytoplasm, which is colourless to pale green. 



Note: A method of counting the cells is given in the original 

 paper. 



Reference: Briseno-Castrejon, B. & Finerty, J. C. (1949). 



ALLOXAN - NINHYDRIN - SCHIFF - LUXOL FAST BLUE 



(W. M. Shanklin & M. Issidorides, i960) 



Solutions required: 



A. Ninhydrin 0-5% or alloxan 1% 

 in absolute alcohol 



143 



