MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



VICTORIA BLUE - KERNECHTROT - LIGHT GREEN 



For elementary bodies of various viruses, notably those of 

 canary pox, ectromelia, variola, and to a lesser degree, 



varicella and herpes simplex 



Solutions required: 



A. Kernechtrot (Herzberg) .. .. o-i gm. 



Aluminium sulphate . . • . 5 gm. 



Distilled water . . . . . . loo ml. 



Dissolve by boiling ; allow to cool ; then filter 

 and add: 



Acetic acid io% aqueous . . . . 0*5 ml. 



B. Light green 1% aqueous . . . . 100 ml. 

 Acetic acid 10% . . . . . . 0-5 ml. 



C. Tartaric acid 10% aqueous. 



D. Victoria blue 4R 1% aqueous . . 10 ml. 

 Distilled water . . . , . . 90 ml. 

 Acetic acid 10% aqueous . . . . 5 ml. 



Technique: 



1 . Make smears of infected tissue on scrupulously clean slides ; 

 allow to dry in the air. 



2. Fix by flaming gently. 



3. Wash in distilled water for ten minutes. 



4. Stain in the Kernechtrot solution for an hour. 



5. Rinse for thirty seconds each in two changes of distilled 

 water. 



6. Drain and allow to dry for half an hour in the atmosphere. 



7. Stain in the light green solution for two minutes. 



8. Rinse for five seconds in each of two changes of distilled 

 water. 



9. Immerse in the tartaric acid solution for two minutes. 



10. Drain oflp the excess tartaric acid solution; then wash for 

 five to ten seconds in each of two changes of distilled water. 



11. Stain with the victoria blue solution for two minutes. 



404 



\ 



