MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



METHYL VIOLET - METANIL YELLOW 

 For typhus fever rickettsiae in lungs of mice 



Solutions required: 



A. Methyl violet loB . . . . . . o-oi% aqueous. 



B. Distilled water . . . . . . 90 ml. 



Acetic acid I % .. .. .. 10 ml. 



C. Metanil yellow . . . . . . o-oi% aqueous. 



Technique : 



1. Fix pieces of lung in 10% neutral formalin. 



2. Dehydrate, clear and embed in paraffin wax in the usual 

 manner. 



3. Fix sections to slides and take down to distilled water as 

 usual. 



4. Stain in the methyl violet solution for half to one hour. 



5. Differentiate in solution B, controlling by examination under 

 the microscope until the cytoplasm is decolorized. 



6. Counterstain in the metanil yellow solution for a few seconds. 



7. Dehydrate in acetone. 



8. Mount in D.P.X. or Clearmount. 



Results: 



Rickettsiae are stained violet. 



Reference: Nyka, W. J. (1945), J^. Path, and Bad., 52, 317-24. 



METHYL VIOLET - PYRONIN - ORANGE G (Bonney's 



Triple Stain) 



For chromatin, connective tissue, keratin 



Solutions required: 



A. Methyl violet 6B (Jensen) 1% 



aqueous . . . . . . 25 ml. 



Pyronin 10% aqueous . . . . 10 ml. 



Distilled water . . . . . • 65 ml. 



160 



