STAINING, PRACTICAL AND THEORETICAL 



Results: 



Nuclei, blue; melanin and haemosiderin remain unstained in 

 their natural brown colours : haemofuscin, bright red. 



References : 



Lillie, R. D. (1948), p. 130. 

 Mallory, F. B. (1938). 



HAEMATOXYLIN - BIEBRICH SCARLET - 

 PICRO ANILINE BLUE 



For differential staining of connective tissue and muscle 



Solutions required: 



A. Haematoxylin (Weigert) A. 



B. Haematoxylin (Weigert) B. 



C. Biebrich scarlet, 0-2% aqueous 100 ml. 

 Glacial acetic acid . . . . i ml. 



D. Picric acid, saturated aqueous . . 100 ml. 

 Aniline blue, water soluble . . o-i gm. 



E. Acetic acid 1% aqueous. 



Technique: 



1. Tissues should be fixed in 10% formalin and paraffin sections 

 employed. 



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

 of equal parts of Weigert's Haematoxylin A and B. 



3. Wash in tap water. 



4. Stain for three to five minutes in the acetic Ponceau, S 

 (Solution C). 



5. Rinse in distilled water. 



6. Stain for three to five minutes in the picro aniline blue 

 (Solution D). 



7. Wash for three or four minutes in 1% acetic acid solution. 



8. Dehydrate in ascending strengths of alcohol and clear in 

 xylol in the usual manner. 



9. Mount in acid balsam. 



Results: 



Connective tissue, glomerular basement membrane and reti- 

 culum: blue. Muscle and plasma: pink. Erythrocytes: bright red. 

 Reference: Lillie, R. D. (1940). 



232 



