SECTION TWO 



4. Immerse the sections in the stain (solution B) for two to 

 three minutes, with gentle agitation. 



5. Differentiate by agitating gently in 85% ethylene glycol. 



6. Transfer to distilled water for three to five minutes. 



7. Counterstain with Ehrlich or Delafield haematoxylin. 



8. Wash well in tap water: mount in Aquamount. 



Results : 



Nuclei: blue. Fat: orange to red. Cholesterol: red. Normal 

 myelin : unstained. Fatty acids : unstained. 



Reference: Chiffelle, T. L. & Putt, F. A. (1951)- 



SCARLET R 

 For staining fat, etc. in animal tissues 



Solutions required: 



A. Scarlet Red, saturated in equal volumes 



of acetone and 70% alcohol. 



B. Ehrlich or Delafield haematoxyhn. 



Technique: 



1. Tissues are fixed in formalin and frozen sections are em- 

 ployed. 



2. Sections are immersed for a second in 70% alcohol; then 

 stained for two to five minutes, in the Scarlet R solution. 



3. Wash quickly in 70% alcohol, and transfer to distilled water. 



4. Counterstain with Ehrlich or Delafield haematoxylin. 



5. Wash well in tap water; mount in glycerine or Aquamount. 



Results: 



Nuclei: blue. Fat: orange to red. Cholesterol: red. Normal 

 myelin: unstained. Fatty acids: unstained. 



Note: Acetone increases the solubility of the dye, but Chiffele 

 & Putt's method (above) is to be preferred as acetone-alcohol is 

 liable to dissolve out some lipid material from the sections. 



Reference: Michaelis, L. (1901). 



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