158 Connective Tissue (chap. 13) 



Churg and Prado (1956) Method 



fixation: mercuric chloride type excellent. Improve formalin fixation 

 by treating slides overnight in saturated aqueous HgCl2. 



solutions: 



Crystal violet stain: 



crystal violet, C.I. 42555 0.1 gm. 



distilled water 100.0 ml. 



glacial acetic acid 0.2 ml. 



Acidic water: 



glacial acetic acid 0.2 ml. 



distilled water 100.0 ml. 



Phosphomolybdic acid: 



phosphomolybdic acid 1.0 gm. 



distilled water 100.0 ml. 



Chromotrope-aniline blue stain: 



chromotrope 2R, C.I. 16570 I.O gm. 



aniline blue WS, C.I. 42780 0.25 gm. 



HCl, 0.02N (1.7 ml. cone. HCl in 998.3 ml. dis- 

 tilled water) 50.0 ml. 



Dissolve aniline blue in acid with gentle heat. Cool and add chromo 

 trope. Keeps well for months. 



o* 



procedure: 



1. Deparaffinize and hydrate slides to water. Remove HgCl2. 



2. Stain in crystal violet: 2-3 minutes. 



3. Differentiate in acidic water: 3-45 seconds. 



4. Rinse well in distilled water. 



5. Treat with phosphomolybdic acid: 3-45 seconds. 



6. Rinse in distilled water. 



7. Stain in chromotrope-aniline blue: 2 minutes. 



8. Rinse in distilled water. 



9. Dehydrate quickly through 70% and 2 changes 95% alcohol. 

 10. Dehydrate, absolute alcohol, clear and mount. 



results: 



Formalin-fixed tissue without HgCl2'. 



nuclei — purplish 



cytoplasm — pale blue to pink 



