SECTION TWO 



Results: 



Oxidase granules are stained blue while nuclei are pink. Some- 

 times fat is stained also. 



NAPHTHOL BLUE BLACK - HAEMATOXYLIN 

 BRILLIANT PURPURIN - AZOFUCHSIN 



For collagen, reticulum, smooth muscle, etc. 



Solutions required: 



A. Weigert's Haematoxylin A. 



B. Weigert's Haematoxylin B. 



C. Brilliant purpurin R. 



in I % acetic acid .. ., ,. 30 ml. 



Azofuchsin 1% in 1% acetic acid. . 20 ml, 



D. Naphthol blue black . . . . i gm. 

 Picric acid, satd., aqueous . . 100 ml. 



Technique : 



1. Paraffin sections are fixed to slides and taken down to 70% 

 alcohol in the usual manner. 



2. Stain for six minutes in a freshly prepared mixture consisting 

 of equal parts of solutions of A and B. 



3. Wash in tap water. 



4. Counterstain for five minutes in solution C. 



5. Wash in 1% acetic acid. 



6. Stain in solution D for five minutes. 



7. Rinse in 1% acetic acid for two minutes. 



8. Pass through the usual ascending grades of alcohol dehydrate 

 in absolute. 



9. Clear in Xylol. 



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



Results: 



Collagen, reticulum and basement membranes: dark green. 

 Smooth muscle: brown. Nuclei: brownish black. 



Reference: Lillie, R. D. (1945). 



