STAINING, PRACTICAL AND THEORETICAL 



HAEMATOXYLIN - AZOPHLOXINE 

 For muscle, connective tissue, ganglion cells, etc. 



Solutions required: 



A. Lavdowsky's Fixative: 



Formalin (40% formaldehyde) . . 10 ml. 



Glacial acetic acid . . . , . . 2 ml. 



Alcohol 95% . . . . • • 50 ml. 



Distilled water . . . . . . 40 ml. 



B. Haematoxylin (Delafield, Harris or 

 Ehrlich) 



C. Azophloxine . . . . . . 0-2 gm. 



Acetic acid 0-2% . . . . . . 100 ml. 



Add about i ml. chloroform as a preservative. 



D. Acetic acid 0-2% aqueous. 



E. Orange G. . . . . . . . . 2 gm. 



Phosphotungstic acid . . • • 4 gm. 



Distilled water . . . . . . 100 ml. 



Fast Green FCF 0-2 gm. 



Technique: 



1. Fix material in Lavdowsky's mixture or in 10% formalin 

 and embed in paraffin wax. 



2. Stain in Harris, Ehrlich or Delafield Haematoxylin for five 

 to ten minutes. 



3. Blue in tap water or 1% lithium carbonate solution. 



4. Stain in the azophloxine solution for two minutes. 



5. Rinse in 0-2% acetic acid. 



6. Stain in the orange G-fast green solution for one minute. 



7. Differentiate for about five minutes with 0-2% acetic acid. 



8. Rinse in distilled water. 



9. Remove excess distilled water by draining and blotting round 

 the edges of the sections carefully, but do not allow to dry com- 

 pletely. 



ID. Dehydrate directly with two or three changes of absolute 

 alcohol, or with cellosolve. 



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