MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



JOHANSEN'S QUINTUPLE STAIN 



For plant tissue composed of a variety of cell types, such as 



leaves, roots, steins and ovaries 



Solutions required: 



A. Safranin O, i% in cellosolve . . 50 ml. 

 Alcohol 95% . . . . . . 25 ml. 



Sodium acetate 4% . . . . 25 ml. 



Formalin . . . . . . . . i ml. 



B. Crystal violet 1% aqueous. 



C. Absolute alcohol 

 Cellosolve 

 Tertiary butyl alcohol 



D. Fast Green, FCF. . 

 Malachite Green . . 

 Cellosolve 



Dissolve; then add: 

 Tertiary butyl alcohol 

 Absolute alcohol 

 Glacial acetic acid 



E. Orange 2 . . 

 Cellosolve 



Dissolve by warming gently, 

 Clove oil . . 

 Absolute alcohol 

 Tertiary butyl alcohol 



F. Terpineol, extra pure 

 Absolute alcohol 



Tertiary butyl alcohol . . I equal 

 Cellosolve . . . . f volumes 



Methyl salicylate 

 Beechwood creosote 



25 ml. 

 25 ml. 

 25 ml. 



0-5 gm. 

 0-5 gm. 

 100 ml. 



25 ml. 

 25 ml. 

 1*5 ml. 



0-5 gm. 

 58 ml. 



then add : 

 14 ml. 

 14 ml. 

 14 ml. 



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