SECTION TWO 



F. Chrome alum, 3% 



G. Potassium dichromate, 5% 



H. Chrome alum haematoxylin 



Solution E (haematoxylin) . . 5 ml. 

 Distilled water . . . . • • 45 nil- 

 Solution F (chrome alum) . . 50 ml. 

 Solution A (sulphuric acid) . . i ml. 

 Solution G (potass, dichromate) 2 ml. 



Note: Solution H will keep for several weeks with- 

 out deterioration. 



I. Alcohol, 70% . . . . • • 99 nil. 



HCl cone. . . . . . . I ml. 



J. Biebrich scarlet, 1% aqueous 



Technique: 



1. Fix pieces of tissue in 10% neutral formalin, and embed in 

 parafRn wax or in celloidin, or cut frozen sections. 



2. Take sections through to water. 



3. Oxidize in solution C for one to two minutes. 



4. Bleach in solution D for one minute. 



5. Wash well in running water. 



6. Stain in solution H for ten minutes. 



7. Differentiate in acid alcohol (solution I). 



8. Wash and " blue " in tap water, or in 1% lithium carbonate 

 solution. 



9. Wash in distilled water. 



ID. Counterstain in the Biebrich scarlet solution. 



1 1 . Wash in running water. 



12. Dehydrate through the usual graded alcohols. 



13. Clear in xylol or benzene. 



14. Mount in D.P.X. or Clearmount or Canada balsam in 

 xylol. 



Results: 



Lipofuscins: dark blue. Nuclei: purple. Cytoplasmic struc- 

 tures: scarlet. 



Reference: Gomori, G. (1952). 



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