STAINING, PRACTICAL AND THEORETICAL 



Technique: 



1. Fix material in io% formalin or Bouin or calcium-formol. 



2. Cut frozen sections, and drop them into solution C in a 

 watch glass, or in a small petri dish. 



3. Cover the watch glass or petri dish to prevent evaporation 

 of the alcohol. 



4. Allow the stain to act from one to ten minutes. 



5. Differentiate, if necessary, in solution D. 



6. Wash well in tap water. 



7. Immerse the sections in solution E for about one minute if 

 it is desired to retain the stain in permanent preparations. 



8. Rinse well in distilled water. 



9. Mount in Aquamount. 



10. If desired, seal the edges of the coverslips with Laktoseal. 



Notes: 



(a) Any of the oil-soluble azo dyes (oil red O, oil red 4B, 

 Sudan red 7B, Sudan black, Sudan blue, Sudan green, etc.) may 

 be used. 



(b) Crystal violet or a number of other basic dyes used as 

 nuclear stains, or haemalum, can be used instead of toluidine 

 blue. 



(c) Lipids are distinctly coloured with the oil-soluble dye. 

 Toluidine blue demonstrates labrocytes and other metachromatic 

 elements; while crystal violet, used in place of toluidine blue in 

 the above technique, is said to show amyloid degeneration very 

 well. 



(d) I would suggest that ethylene or propylene glycol might be 

 used to advantage in place of isopropyl alcohol, in cases where more 

 intense staining of lipids is desired as more concentrated solutions 

 of the oil-soluble dyes can be obtained with this solvent, which 

 like isopropyl alcohol, is miscible with water. 



Reference: Arvy, Lucie (1958). 



