SECTION TWO 



Dissolve; then add: 



Zinc metal powder . . . . lo gm. 



Glacial acetic acid . . . . 2 ml. 



Boil until the blue colour completely disappears. 

 Allow to cool ; shake with about i gm. of decolorizing 

 carbon; then filter. The liquid which should then 

 be quite colourless is stored in a well stoppered 

 bottle. 



B. Solution B . . . . . . 10 ml. 



Glacial acetic acid . . . . 2 ml. 



Hydrogen peroxide 3% . . i ml. 



N.B. : This solution must be freshly prepared and 

 filtered before use. 



C. Safranin o-i% aqueous . . 99 ml. 

 Glacial acetic acid . . . . i ml. 



Technique: 



1. Fix tissue blocks, not more than 3 to 5 mm. in thickness in 

 10% formalin buffered to pH 7-0 for 24 to 28 hours (prolonged 

 fixation should be avoided). 



2. Embed in paraffin wax as usual and cut section 5 to 6/x in 

 thickness. 



3. Take sections down to water as usual. 



4. Stain in solution B for three to five minutes. 



5. Wash briefly in water. 



6. Counterstain for thirty to sixty sections in the safranin solu- 

 tion. 



7. Rinse briefly with water. 



8. Dehydrate as usual. 



9. Clear in xylol. 



10. Mount in Clarite or other synthetic mountant such as 

 D.P.X., Clearmount, etc. 



Results: 



Haemoglobin, dark blue-green ; background light pink. 



Note: Blood and tissue smears fixed with methyl alcohol may 

 also be stained by applying the stains as prescribed above. 



Reference: Adapted from Dunn, R. C. (1946), Stain. Tech., 21, 65. 



