MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



B. Absolute alcohol . . . . . . 30 ml. 



Phosphate buffer 1/15 M (pH 7-2). . 10 ml. 



Distilled water . . . . . . 20 ml. 



Tetrazotized o-anisidine . . . . o-o6 gm. 



Technique: 



1. Tissues are fixed in 10% formalin (or 5% formol saline). 



2. Without washing out fixative, cut frozen sections and place 

 them on slides. 



3. Allow the sections to become affixed to slides by drying in 

 the atmosphere; then wash out formalin with several changes of 

 water. 



4. Immerse in solution B for one or more hours in an incubator 

 at room temperature. 



5. Remove excess reagent (solution B) by washing for two hours 

 in 50% alcohol (aldehyde free). 



6. Wash in several changes of water for several hours. 



7. Immerse in solution C for one to two minutes. 



8. Wash in several changes of distilled water to which a few 

 drops of acetic acid have been added. 



9. Drain off excess liquid; then mount in Glycerin Jelly. 



Results: 



Sites of carbonyl activity are indicated by the production of a 

 blue pigment. 



Reference: Seligman, A. M. and Ashbel, R. (195 1), Cancer, 4, 579-96. 



METANIL YELLOW - IRON HAEMATOXYLIN 



For radioautographs prepared by mounting tissue sections 

 directly onto photographic emulsions for study of thyroid 

 carcinoma and all organs of man, as well as laboratory 

 animals to which radio isotopes have been administered 



Solutions required: 

 A. Metanil Yellow 0-25% aqueous. 



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