134 Hematoxylin Staining (chap. 12) 



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



3. Wash in running water: 3 minutes. 



4. Blue in Scott's solution: 3 minutes. 



5. Wash in running water: 3-5 minutes. 



6. Counterstain. 



7. Dehydrate quickly through 70 and 95% alcohols. 



8. Finish dehydration, absolute alcohol. 



9. Clear and mount. 



results: 



nuclei — deep blue 



other elements — colors of counterstain 



Heidenhain's Iron Hematoxylin 



fixation: any general fixative, preferably one containing HgClo. 



solutions: 



Hematoxylin and iron alum, page 127. 

 Counterstains, page 129. 



procedure: 



1. Deparaffinize and run slides down (hydrate) to water, removing 

 HgCl2. 



2. Mordant in 4% iron alum: 15-30 minutes (or overnight, see com- 

 ments below). 



3. Wash in running water: 5 minutes. 



4. Stain in hematoxylin: 15-30 minutes (or overnight). 



5. Wash in running water: 5 minutes. 



6. Destain in 2% aqueous iron alum or saturated aqueous picric acid 

 (see comments below) until nuclei stand out sharp and clear against 

 colorless background. Dip slides in water containing couple of 

 drops of ammonia before examining them. Warning: too much 

 ammonia will loosen sections. 



7. Wash in running water: 15-30 minutes or longer. 



8. Counterstain, if desired. 



9. Run slides up through alcohols (dehydrate), clear and mount. 



results: 



nuclei — blue black to black 



muscle striations — blue-grey \ 



other elements — colors of counterstain, or very light grey to colorless 



if no counterstain used \ 



