100 STAINING METHODS 



rub up in a mortar with 



Distilled water 100.0 c.c. 



Add 



Fuchsin, saturated alcoholic solution ... 10.0 c.c. 



and make up to 200 c.c. by the addition of 



Distilled water 90.0 c.c. 



Filter. 



Allow the stain to stand for two weeks before use; 

 keep in a dark place or in an amber glass bottle. Owing 

 to the unstable character of the methyl green, this 

 stain deteriorates after about six months. 



METHOD. 



1. Prepare and fix film in the usual manner. 



2. Flood the cover-slip with the stain and allow it 

 to act for five to ten minutes. 



3. Wash very thoroughly in water;' if necessary, 

 direct a powerful stream of water on the film from a 

 wash-bottle. 



4. Dry and mount. 



2. Muir's Method. 



1. Prepare, dry and fix film in the ordinary manner. 



2. Flood the film with carbolic fuchsin, warm until steam 

 begins to rise. Allow the stain to act for thirty seconds. 



3. Wash quickly with methylated spirit. 



4. Wash thoroughly with water. 



5. Subject the film to the action of the following mordant for 

 five seconds: 



Corrosive sublimate, saturated aqueous solution . 2 c.c. 

 Tannic acid, 20 per cent, aqueous solution ... 2 c.c. 

 Potash alum saturated aqueous solution . . . . 5 c.c. 



6. Wash thoroughly in water. 



7. Treat with methylated spirit for about sixty seconds. (The 

 preparation should now be pale red.) 



8. Wash thoroughly in water. 



9. Counterstain in methylene blue, aqueous solution thirty 

 seconds. 



10. Wash in water. 



11. Dehydrate in alcohol. 



1 2 . Clear in xylol and mount in xylol balsam. 



