STAINING, PRACTICAL AND THEORETICAL 



B. Lugol's iodine 



C. Sodium thiosulphate 0*5% aqueous 



D. Delafield Haematoxylin 



E. Alcohol 95% . , . . . . 999 ml. 

 Aniline oil . . . , . . . . i ml. 



F. Azocarmine G or B 1% aqueous . . 100 ml. 

 Glacial acetic acid . . . . . . 4 ml. 



G. Glacial acetic acid . . . . . . 10 ml. 



Alcohol 90% . . . . . . 90 ml. 



H. Phosphotungstic acid 5% 



1. Acid green L extra . . . . . . o-i gm. 



Orange G . . . . . . . . 0-5 gm. 



Clove oil . . . . . . . . 100 ml. 



N.B. Use fresh stain for each batch of about twenty slides. 



Technique : 



1. Fix in solution A for six to twelve hours. 



2. Wash six to twelve hours in running water. 



3. Dehydrate by immersing for thirty minutes in each of the 

 following: 30%, 50%, 70%, 80% and 95% alcohol. 



4. Immerse in absolute alcohol, two changes, for one hour in 

 each. 



5. Immerse in a mixture consisting of equal parts of absolute 

 alcohol and cedarwood oil for one hour. 



6. Cedarwood oil, one to sixteen hours. 



7. Xylol, for fifteen minutes. 



8. Infiltrate in paraffin wax 56-58° C. (four changes before 

 finally embedding). 



9. Fix sections, 4ju, in thickness, to slides and remove paraffin 

 wax by immersing in two changes of xylol for three minutes each. 



10. Immerse for three minutes in each of two changes of 

 absolute alcohol. 



11. In 95% alcohol for three minutes. 



12. Distilled water for three minutes. 



13. Lugol's Iodine for three minutes. 



14. Sodium thiosulphate solution for three minutes when the 

 sections should have been restored to their natural colour. 



142 



