My elm 21. '5 



Periodic acid, 0.5%: 



periodic acid 0.5 gm. 



distilled water 100.0 ml. 



Lithium carbonate, 0.05%: 



lithium carbonate 0.5 gm. 



distilled water 1000.0 ml. 



Schiff's reagent, see page 294. 



SulfuroLis acid: 



sodium metabisulfite, 10% aqueous 6.0 ml. 



distilled water 100.0 ml. 



IN HCl (see page 408) 5.0 ml. 



Mayer's hematoxylin, see page 125. 



procedure: 



1. Deparaffinize and run slides down to 95% alcohol, but remove 

 HgCl2 if present. 



2. Stain in Luxol fast blue, 60°C: overnight. 



3. Rinse off excess stain in 95% alcohol. 



4. Rinse in distilled water. 



5. Dip in lithium carbonate: 15 seconds. 



6. Differentiate in 70% alcohol: 20-30 seconds. 



7. Rinse in distilled water. 



8. Place in lithium carbonate, second solution: 20-30 seconds. 



9. Differentiate in 70% alcohol: 20-30 seconds. 



10. Rinse in distilled water. If differentiation is not complete, repeat 

 steps 8 and 9. 



1 1. Oxidize with periodic acid: 5 minutes. 



12. Wash in distilled water, 2 changes: 5 mintites. 



13. Treat with Schiff's reagent: 15-30 minutes. 



14. Treat with sulfurous acid, 3 changes: 2 minutes each. 



15. Running water: 5 mintites. 



16. Stain in Mayer's hematoxylin (or the like): 3 minutes. 



17. Wash in running water: 5 minutes. 



18. Dehydrate, clear, and mount. 



results: 



myelin — blue-green 



PAS positive elements — rose to red 



nuclei — blue 



