SECTION SIX 



8. Differentiate in 5% acetic acid for thirty minutes. 



9. Rinse thoroughly in distilled water. 



10. Refine the differentiation in the 0-5% HCl (Solution D). 



1 1 . Wash in several changes of distilled water. 



12. Mount in glycerine jelly. 



Results: 



Phospholipids, blue. Nuclei, red. 



Reference: Menschik, Z. (1953), Stain Tech., 28, 13-18. 



PHOSPHOMOLYBDIC ACID - EOSIN 



For the histochemical localization of choline-containing 



lipids, in frozen sections 



Solutions required: 



A. Acetone . . . . . . . . i volume 



Ether . . * . . . . . i volume 



B. Chloroform . . . . . . . . i volume 



Absolute alcohol , . . . . . i volume 



C. Phosphomolybdic acid. 

 I % in solution B. 



D. Stannous chloride 1%. 

 in 3N hydrochloric acid. 



E. Eosin 1% aqueous. 



Technique: 



1. Dip frozen sections into acetone-ether. 



2. Immerse in the phosphomolybdic acid solution for fifteen 

 minutes. 



3. Rinse in solution B. 



4. Dip into solution D. 



5. Counterstain with the eosin solution for one to two minutes. 



6. Mount in glycerine jelly. 



Results: 



Positive areas are stained blue, whilst negative areas are red. 



Reference: Landing, B. H. (1952), Uzman, L. L. and Whipple, Ann. Lab. 

 Invest., I, 456-2. 



