2?54 Pigments mid Minerals (chap, 17) 



procedure: 



1. Deparaffinizc and hydrate slides to water; remove HgCli.. 



2. Wash in distilled water: 1-2 min4.ites. 



3. Place slides in equal parts of hydrochloric acid and potassium 

 ferrocyanide mixed immediately before use: 20 minutes. Do not 

 handle slides with metal forceps. 



4. Wash thoroughly in distilled water {not tap water). 



5. Coiuiterstain with safranin (or other red nuclear stain): 2-3 

 minutes. 



6. Rinse in 70% alcohol. 



7. Dehydrate, clear, and mount. 



results: 



iron pigment — brilliant greenish blue 



nuclei and other tissue elements — shades of rose and red 



comments: 



For masked or occult iron (Glick, 1949), pre-treat slides with 30% 

 hydrogen peroxide alkalized with dilute ammonia (1 drop/100 ml. 

 peroxide): 10-15 minutes. Wash well and proceed to step 3. If the 

 hydrogen peroxide tends to form bubbles under the sections and 

 loosen them, keep the solution and slides cool in the refrigerator dur- 

 ing the treatment. 



Iron Reaction (hutchison, 1953) 



fixation: Hutchison recommends: 



sodium sulfate 12.0 gm. 



glacial acetic acid 33.0 ml. 



formalin 40.0 ml. 



distilled water to make 200.0 ml. 



Go directly into 70% alcohol from fixative. 



solutions: 



Ferrocyanide-hydrochloric acid solution: 



{a) potassium ferrocyanide 2.0 gm. 



distilled water 50.0 ml. 



(h) hydrochloric acid 2.0 ml. 



distilled water 50.0 ml. 



Freshly mix above solutions, warm slightly and filter. Place in 

 paraffin oven, 56°C a short tiine before using. 



Safranin, see page 233. 



