Pituitary and Fayicrcas 285 



Mayer's hematoxylin, see page 125. 



Orange G Solution: 



orange G, C.I. 162-)0 2.0 gm. 



phosphotungstic acid, 5% (5 gm./lOO ml. water) 100.0 ml. 



Allow to stand 48 hours. Use supernatant fluid. 



procedure: 



1. Deparaffinize and hydrate slides to water; remove HgCU. 



2. Transfer to 70% alcohol: 2 minutes. 



3. Oxidize in periodic acid: 5 minutes. 



4. Rinse in 70% alcohol. 



5. Place in reducing solution: 1 minute. 



6. Rinse in 70% alcohol. 



7. Treat with Schiff's reagent: 10-30 minutes. 



8. Treat wath sulfite solutions, 3 changes: 1 minute, 2 minutes, and 

 2 minutes. 



9. Wash in running water: 10 minutes. 



10. Stain in celestin blue: 2 minutes. 



11. Stain in Mayer's hematoxylin: 2 minutes. 



12. Wash in running water: 5 minutes. 



13. Stain in orange G: 10 seconds. 



14. W^ash in rtinning water until orange color is differentiated: about 

 10-15 seconds. Check under microscope. 



15. Dehydrate, clear, and mount. 



results: 



colloid of stalk and parenchyma, and vesicles of vesiculate chromo- 

 phobes — magenta 

 basophil {beta) granules- — dark red 

 acidophil (alpha) granules — orange 

 erythrocytes — orange 

 nuclei — dark blue 



Trichrome PAS (lazarus, 1958) 



fixation: Zenker-formol (Helly's). Bouin's unsatisfactory. 



solutions: 



Schiff's reagent, see page 294. 

 Sulfurous acid: 



sodium metabisulfite, Na^S^Or, 0.5 gm. 



distilled water 100.0 ml. 



