SECTION TWO 



to the stain solution ; then make up the final volume 

 to 150 ml. with distilled water. 



Note: Solution H deteriorates after repeated use, 

 and it is recommended that the solution should be 

 renewed after each batch of sections. 



I. Lissamine fast yellow . . . . o-i gm. 



Acetic acid, 1% aqueous . . loo ml. 



or 



J. Light green, 0-25% aqueous 



Technique: 



1. Fix material in Bouin, or formol-saline or formol-sublimate, 

 or Zenker (acetic or formol), or alcohol, or formol-acetic-alcohol. 



2. If a mercury-containing fixative has been used, treat for the 

 removal of mercuric precipitate. 



3. Stain sections with the celestin blue solution for two minutes. 



4. Rinse in tap water. 



5. Stain in the haemalum solution for two minutes. 



6. Rinse and blue in the lithium carbonate solution. 



7. Differentiate in the acid alcohol (solution D), controlling by 

 microscopic examination, until the cytoplasm is clear and the 

 nuclei are delicately but firmly revealed. 



8. Immerse in Barlow's tribasic stain (solution H) for five 

 to fifteen minutes. 



9. Wash quickly in running water. 



ID, Counterstain lightly with solutions I or J. 



1 1 . Rinse in water. 



12. Dehydrate quickly with alcohol; clear in xylol; and mount 

 in D.P.X., Cristalite, Clearmount, or Emexel. 



Results: 



Mast-cell granules are stained orange to red, the shade varying 

 somewhat with the species of animal. Cartilage, keratin, and 

 bacteria: pink or red. Mucin is occasionally stained a faint pink. 

 Nuclei are blue, and other tissue elements, green or clear yellow, 

 depending on which of the two counterstains (solutions I and J) 

 have been employed. 



The method gives satisfactory results on the tissues of man, ox, 

 pig, dog, and rat. 



449 



