STAINING, PRACTICAL AND THEORETICAL 



Technique: 



1. Fix pieces of lung in io% 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). 



METHYL VIOLET - PYRONIN - ORANGE G 

 (Bonney's Triple Stain) 



For chromatin, connective tissue, keratin 



Solutions required: 



A. Methyl violet 6B (Jensen) 1% 



aqueous 

 Pyronin B io% aqueous. 

 Distilled water 



B. Acetone 

 Orange G aqueous 2% 



Add orange G solution drop by drop to the ace- 

 tone, with shaking, until the flocculent precipitate 

 formed just redissolves with further addition of 

 orange G solution. 



Technique: 



1. Small pieces of tissue are fixed in acetic-alcohol or in mer- 

 curic chloride. 



2. Wash; dehydrate; clear; embed in paraffin wax. 



25 ml. 

 10 ml. 

 65 ml. 



100 ml. 

 About 10 ml. 



300 



