SECTION TWO 



7. Mordant in solution C (iron alum) for one hour. 



8. Rinse well in tap water. 



9. Wash in distilled water. 



10. Immerse in the haematoxylin (solution E) for two to twenty- 

 four hours. 



11. Dehydrate in two changes of absolute isopropyl alcohol. 



12. Clear in xylol. 



13. Mount in D.P.X. or Emexel or Canada balsam in xylol. 



Notes: 



(a) The authoress states that despite the number of mordants 

 proposed, it is difficult to make consistently good phosphotungstic 

 acid haematoxylin preparations of formalin-fixed material. The 

 difficulty may be due to inadequately oxidized staining solution, 

 but the method described above has been found to give results 

 that have been completely successful in every detail. 



(b) Staining in the haematoxylin solution is usually complete 

 in two or three hours, and should be checked at intervals, by 

 microscopic examination. 



(c) Bleaching in oxalic acid and potassium permanganate is 

 stated to be necessary to give sharp contrast. 



Reference: Lieb, E. (1948). 



PHOSPHOMOLYBDIC ACID - EOSIN 

 For choline-containing lipids ' 



Solutions required: 



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



Ether . . . . . . . . i volume 



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



Absolute alcohol , . . . . . i volume 



C. Phosphomolybdic acid. 

 I % in solutionlB. 



D. Stannous chloride 1%. 

 in 3N hydrochloric acid. 



E. Eosin 1% aqueous. 



Technique: 



I . Dip frozen sections into acetone-ether. 



N 



373 



