138 Hematoxylin Staining (chap. 12) 



procedure: 



1. Deparaffinize and hydrate slides to water, remove HgCIa- 



2. Stain in Weigert's hematoxylin: 3-5 minutes or longer. 



3. Wash, running water: 5 minutes. 



4. Counterstain. 



5. Dehydrate, clear, and mount. 



results: 



nuclei — black 



other elements — color of counterstain 



comments: This is used progressively, check results under microscope. 



Mallory's Iron Chloride Hematoxylin (1944) 

 fixation: any general fixative, or one specific for nuclei. 



solutions: 



Ferric chloride, page 128. 

 Hematoxylin, page 128. 

 Counterstains, page 129. 



Ferric chloride for destaining (0.25%): dilute 5 ml. of the 5% stock 

 solution (above) with 95 ml. distilled water. 



procedure: 



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



2. Mordant in ferric chloride: 1 hour or longer. 



3. Wash in running water: 5 minutes. 



4. Stain in hematoxylin: 1 hour or longer. 



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



6. Destain in ferric chloride, 0.25%; agitate slides, check under 

 microscope. 



7. Wash thoroughly, running water: 10-15 minutes. 



8. Counterstain, if desired. 



9. Dehydrate, clear, and mount. 



results: 



nuclei — deep blue 



other elements — color of counterstain 



Hematoxylin Substitute Procedures 



Gallocyanin (romeis, 1948) 



Gallocyanin is an excellent substitute for hematoxylin and can be 

 used in most procedures designed for the latter dye. The solution is 



