136 Hematoxylin Staining (chap. 12) 



procedure: 



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



HgCl2. 



2. Stain in Groat's iron hematoxylin: approximately 10 minutes. 



3. Rinse off excess stain with running water: 10 minutes. 



4. Counterstain, if desired. 



5. Dehydrate, clear, and mount. 



results: 



nuclei — black 



other elements — color of counterstain 



comments: 



The 10-minute timing is approximate: check under the microscope 

 for depth of color. If the background is too grey it may be decolorized 

 in: 



distilled water 50.0 ml. 



95% alcohol 50.0 ml. 



sulfuric acid (sp.gr. 1.84, 95.5-96.57o H2SO4) ... 0.18. ml. 



Follow by neutralizing in: 



distilled water 50.0 ml. 



95% alcohol 50.0 ml. 



sodium bicarbonate 0.5 gm. 



Phosphotungstic Acid Hematoxylin (as modified by lieb 1948) 



fixation: any general fixative, preferably containing HgClo. Lieb sug- 

 gests if tissue is formalin fixed, mordant sections before staining in 

 saturated aqueous mercuric chloride, rinse and treat with Lugol's. 



solutions: 



Hematoxylin, page 125. 

 Potassium permanganate, 0.5% 



potassium permanganate 0.5 gm. 



distilled water 100.0 ml. 



Oxalic acid, 2% 



oxalic acid 2.0 gm. 



distilled water 100.0 ml. 



Iron alum, 4^, page 127. 



