STAINING, PRACTICAL AND THEORETICAL 



B. Schiff reagent 



C. Sodium bisulphite M/i (lOO gm,/ 



litre NaHSOs) 



D. Solution C 

 Tap water 



D. Luxol fast blue (Michrome No. 



331) 



Alcohol 95% 



10 ml. 

 200 ml. 



I gm. 

 I litre 



Technique: 



1. Sections of material that have been fixed in 10% formalin 

 are passed through two changes of xylol. 



2. Pass through two changes of absolute alcohol. 



3. Incubate at 37° C for five to twenty-four hours in ninhydrin 

 or alloxan (solution A.) 



4. Wash gently under the tap for one to two minutes. 



5. Immerse in Schiff 's reagent for one-half to one hour. 



6. Pass sections through three jars of solution D, for ninety 

 seconds in each, with frequent agitation. 



7. Wash in tap water for five minutes. 



8. Dehydrate in 95% alcohol. 



9. Counterstain in the luxol fast blue solution for five minutes. 



10. Pass through absolute alcohol. 



11. Clear in xylol and mount in Canada balsam in xylol or 

 D.P.X. 



Results: 



Human brain: protein material in the nucleus supraopticus of 

 the hypothalamus and in the hypoglossal nucleus is stained red 

 by alloxan- Schiff treatment, while the Nissl bodies are stained 

 blue by the luxol fast blue. 



Notes 



(a) The authors (Shanklin & Issidorides) state that Lillie (1954) 

 in describing Yasuma & Ichikawa's (1953) technique observes 

 that counterstains in his hands had not been found successful so 

 far. In the method described above, however, Shanklin and 

 Issidorides found luxol fast blue highly satisfactory as a counter- 

 stain for the alloxan-Schiff method. 



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