SECTION TWO 



PHLOXIN - METHYLENE BLUE - AZUR B 

 For normal and pathological animal tissues 



Solutions required: 



A. Phloxin . . . . . . , . 0-5 gm. 



Acetic acid 0-2% aqueous . . . . 100 ml. 



Filter before use. 



B. Methylene blue . . . . . . 0-25 gm. 



Azur B . . . . . . . . 0-25 gm. 



Borax. . . . . . . . . . 0-25 gm. 



Distilled water . . . . . . 100 ml. 



C. Acetic acid 0-2% aqueous. 



Technique: 



1 . Fix sections to slides : remove paraffin and pass through the 

 usual descending grades of alcohol to distilled water. 



2. Immerse in solution A for one to two minutes. 



3. Rinse well in water. 



4. Stain for one half to one minute in solution B, 



5. Destain partially in solution C. 



6. Differentiate in three washes of 95% alcohol. 



7. Dehydrate with two changes of absolute alcohol. 



8. Clear in two changes of xylol. 



9. Mount in synthetic medium (such as D.P.X. or Clearmount, 

 etc.). 



Results: 



Nuclei and bacteria are stained blue with some metachromasia. 

 Collagen and other tissue elements are bright pink to red. Erythro- 

 cytes, bright scarlet. 



Note: 



This is a rapid modification of Mallory's original method re- 

 ducing the staining time from one hour or more to one to two 

 minutes, and permitting good staining with formalin fixed tissues 

 vv^hich is not possible with Mallory's original method which was 

 designed for Zenker-fixed material. Colophony differentiation is 

 obviated, and Phloxin is not washed out as in the original 

 Mallory technique. 



Reference: Thomas, John T. (1953). 



