Hemoglobin Staining 239 



3. Drain, and flood with peroxide reagent: 1.5 minutes. 



4. Wash in distilled water: 15 seconds. 



5. Counterstain if desired with a red nuclear stain. 



6. Dehydrate, clear, and mount. 



results: 



hemoglobin — dark browai 



For hemosiderin, see iron reactions, page 233. 



Cyanol Reaction (dunn, 1946) 



fixation: formalin, preferably buffered to pW 7; fix no longer than 48 

 hours. Dried smears may be methyl alcohol fixed and then proceed to 

 step 2 below. 



solutions: 



Cyanol reagent: 

 Stock solution: 



cyanol I.O gm. 



distilled water 100.0 ml. 



zinc powder, C.P 10.0 gm. 



glacial acetic acid 2.0 ml. 



Bring to boiling point. In short time the solution is decolorized. 

 Stable for several w^eeks. 



Working reagent: 



Filter 10 ml. of stock reagent, and add: 



glacial acetic acid 2.0 ml. 



hydrogen peroxide, commercial 3% 1.0 ml. 



procedure: 



1. Deparaffinize and hydrate slides down to water. 



2. Treat with working reagent: 3-5 minutes. 



3. Rinse in distilled water. 



4. Counterstain in red nuclear stain. 



5. Dehydrate, clear, and mount. 



results: 



hemoglobin — dark blue to bluish grey 



comments: 



Gomori (1952) uses 1 gram acid fuchsin instead of cyanol and pro- 

 duces a red hemoglobin. Counterstain with hematoxylin or other 

 blue nuclear stain. 



