GENERAL STAINING 657 



1316. Breinl's Triple Stain (safranin, polychrome methylen blue 

 and orange tannin). Mordant fifteen minutes in alcoholic iodine- 

 potassium iodide solution, rinse with water and stain thirty 

 minutes or longer in safranin (alcoholic solution). Wash with 

 water, place ten minutes in polychrome methylen blue and again 

 wash with water. Flood with orange tannin. When the orange 

 has replaced the blue in the cytoplasm, wash well with 95 per cent, 

 alcohol, rinse in absolute alcohol and clear with anilin oil, which 

 also completes the differentiation. The action is not given by pure 

 anilin oil, but by virtue of some impurity in the commercial 

 product. Rinse with cedar oil, and mount in balsam. Chromo- 

 some tlu-eads blue, metaphase chromosomes red, cytoplasm pale 

 yellow. 



