Trichrome Staining 153 



Masson C: 



fast green FCF, C.I. 42053 2.5 gm. 



distilled water 100.0 ml. 



glacial acetic acid 2.5 ml. 



Aniline blue can be substituted for fast green. If so, after staining 

 with Masson C (step 1 1 below), extract excess stain for 5 minutes with 

 1% phosphomolybdic acid, aqueotis, followed by a quick rinse in 

 distilled water and then proceed with step 12, below. 



Hematoxylin, see page 124. 

 Acetic acid rinse. 



glacial acetic acid 1.0 ml. 



distilled water 100.0 ml. 



procedure: 



1. Deparaffinize and hydrate slides to water. Remove HgCU. Mor- 

 dant formalin-fixed tissue in saturated aqueous HgClo: 5 min- 

 utes. Wash, running water: 5 minutes. Treat with Lugol's and 

 sodium thiosulfate. 



2. Mordant ^vith iron alum: | hour at room temperature or 5 min- 

 utes at 40-50°C. 



3. Wash in running water: 5 minutes. 



4. Stain in hematoxylin (Delafield's or the like): ^ hour, room tem- 

 perature, or 5 minutes at 40-50°C. 



5. Wash in running water: 5 minutes. 



6. Differentiate in saturated aqueous picric acid. 



7. Wash thoroughly, running water: 10-20 minutes. 



8. Stain in Masson A: 5 minutes. 



9. Rinse in distilled water. 



10. Treat with Masson B: 10 minutes. 



11. Stain in Masson C: 2-5 miniues. 



12. Differentiate in acetic acid rinse: 1-2 minutes. 



13. Dehydrate quickly 707^ and 95% alcohol. 



14. Dehydrate, absokue alcohol, 2 changes: 3 mintites each. 



15. Clear and mount. 



results: 



nuclei — blue to blue-black 

 collagen, mucus — blue or green 

 cytoplasmic elements, keratin, muscle — reds 



