MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



MASSON'S TRICHROME STAIN 

 For connective tissues 



Solutions required: 



A. Iron alum 5% aqueous. 



B. Regaud's haematoxylin solution. 



C. Picric acid, saturated in 95% 



alcohol . . . . . . . . 20 ml. 



Alcohol 95% . . . . . . 10 ml. 



D. Ponceau fuchsin. 



E. Phosphomolybdic acid 1% aqueous. 



F. Aniline Blue 5% in 2% acetic acid. 



Technique: 



1. Fix pieces of tissue in Bouin's fluid for three days or in 

 Regaud's fluid for one day. 



2. Wash in running water; dehydrate; clear and embed in 

 parafiin wax as usual. 



3. Sections 5/1 in thickness are fixed to slides; de-waxed and 

 passed through descending grades of alcohol down to distilled 

 water in the usual manner. 



4. Mordant in Solution A for five minutes at 45° C. to 50° C. 



5. Wash well in distilled water. 



6. Stain for five minutes in Regaud's haematoxylin at 45° C. 

 to 50° C. 



7. Rinse in distilled water. 



8. DiflFerentiate in picric alcohol (Solution C above) controlling 

 by examination under the microscope, while the preparation is 

 still wet. 



9. Wash in running tap water for a minute or so. 



10. Stain for five minutes in the Ponceau fuchsin solution. 



11. Rinse in distilled water. 



12. Diff"erentiate in the phosphomolybdic acid solution for five 

 minutes. 



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