SECTION TWO 



Technique: 



1. Fix brains in io% formalin, preferably by perfusion, and 

 sliced to the desired thickness as soon as they are firm enough. 



2. Store the sliced brain in the fixative, for a minimum time of 

 two weeks for human material, or in the case of laboratory animals, 

 for at least one week. 



3. Wash the slices in water for at least six hours. 



4. Dehydrate for one hour in each of two changes of 95% 

 alcohol. 



5. Stain in solution A for sixteen to eighteen hours at 45-55° C. 



6. Remove excess stain by immersing the preparations in 95% 

 alcohol. 



7. Rinse in distilled water. 



8. Differentiate in several changes of solution C. 



9. Refine the differentiation in a number of changes of 70% 

 alcohol. 



10. Rinse in distilled water. 



11. Store in 10% formalin. 



Results: 



Tracts of white matter stain brilliant blue, contrasting strongly 

 with cellular areas of grey matter, stained very pale green. 



Notes: 



The author states that absence of myelin is evident in patho- 

 logical demyelinated regions of adult brain and in the non- 

 myelinated areas of very young animals. 



Reference: Dziabis, M. D. (1958). 



LUXOL FAST BLUE 



For increasing the contrast of sectioned material for phase 



contrast microscopy 



Solutions required: 



A. Luxol fast blue 2% in 95% 



alcohol 



B. Ferric chloride 29% aqueous (FeCls) 



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