286 Cytoplasmic Elements (chap. 18) 



Weigert's hematoxylin, see page 126. 



Ponceau-orange G: 



ponceau 2R, C.I. 16150 0.2 gm. 



orange G, C.I. 16230 0.1 gm. 



distilled water 100.0 ml. 



glacial acetic acid 1.0 ml. 



Light green: 



light green SF yellowish, C.I. 42095 1.0 gm. 



distilled water 100.0 ml. 



glacial acetic acid 1.0 ml. 



Phosphomolybdic acid solutions: 



phosphomolybdic acid 1.0 gm. 



distilled water 100.0 ml. 



To make 0.5% solution, use above 1% 



phosphomolybdic acid 50.0 ml. 



distilled water 50.0 ml. 



procedure: 



1. Deparaffinize and hydrate slides to water: remove HgCl2. 



2. Treat with periodic acid, 0.6% (0.6 gm./lOO ml. water): 20 

 minutes. 



3. Wash in rimning water: 5 minutes. 



4. Treat with Schiff's reagent: 20 miniues. 



5. Transfer through 3 sulfurous acid rinses: 0.5 minute each. 



6. Wash in running water: 5-10 minutes. 



7. Rinse in distilled water. 



8. Stain in Weigert's ferric chloride hematoxylin (24 hours old): 10 

 minutes. 



9. Rinse in 95%, alcohol. 



10. Differentiate in acidic alcohol (0.5 ml. HCl/lOO ml. water). 



11. Wash in running water: 10 minutes. 



12. Stain in ponceau-orange G: 2 hours. 



13. Rinse in distilled water. 



14. Differentiate in phosphomolybdic acid, 1% initil delta cells and 

 collagen are colorless. 



15. Rinse briefly in distilled water. 



16. Stain in light green: 5-30 minutes. 



17. Wash well in acetic acid, 1% (1 ml./99 ml. water): 1-2 minutes. 



18. Differentiate in phosphomolybdic acid, 0.5%: 0.5-5 minutes. 



19. Wash in acetic acid, 1%: 20 miniUes. 



