SECTION FOUR 



SAFRANIN CRYSTAL VIOLET (Hermann) 

 For chromosomes 



Solution required: 



A. Safranin . . . . . . . . i gm. 



Aniline water . . . . • . 50 ml. 



Absolute alcohol . . . . • • 50 ml. 



B. Crystal violet 1% in 70% alcohol. 



Technique: 



1. Stain for three to twenty-four hours in the safranin solution 

 in a closed jar or tube. 



2. Destain briefly with 50% alcohol. 



3. Counterstain with the crystal violet from thirty seconds to 

 five minutes. 



4. Rinse quickly with 90% followed by absolute alcohol. 



5. Clear with clove oil. 



6. Wash with xylol, and mount. 



Results: 



Chromosomes and nucleoli are stained red; resting nuclei and 

 chromatin granules violet: propase and early telophase nuclei 

 may show violet chromonema and red chromatin granules ; cyto- 

 plasm, light violet; spindle, deeper violet. 



Reference: Hermann, Arch, Mik. Anat., 34, 58. 



SAFRANIN - GENTIAN VIOLET - ORANGE G. 

 (Flemming Tricolour stain) 



For chromosomes, etc. 



Solutions required: 



A. Safranin 3*5% in 95% alcohol . . 10 ml. 

 Aniline water . . . . . . 10 ml. 



B. Gentian violet 5% aqueous. 



C. Orange G 1% aqueous. 



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