288 Cytoplasmic Elements (chap. 18) 



5. Rinse in distilled water, 3 changes: 2 minutes each. 



6. Treat with Schiff's reaoent: 20 minutes. 



7. Rinse in tap water. 



8. Stain acidophils in orange G: 10 minutes. 



9. Rinse rapidly in water. 



10. Dehydrate, clear, and mount. 



results: 



thyrotrophs (beta) — deep purple 

 gonadotrophs (delta) — red 

 acidophils, erythrocytes — orange 

 nuclei — unstained 



comments: 



If aldehyde is overstained, treat for aboiu 4 minutes with 0.1% 

 dilution of Clorox. Check under microscope. Wash thoroughly 10 or 

 more minutes. 



If oxidation of tissues is desired before staining, treat with equal 

 parts of 0.2 potassium permanganate and 1% sulfuric acid: 5 minutes. 

 Bleach in 5% oxalic acid. Wash. 



Rhodacyan Method (glenner and lillie, 1957) 



fixation: Formalin or Zenker-formalin, not acetic formalin or Bouin's. 



solution: 



eosin, C.I. 45400, 1% (1 gm./lOO ml. water) ... 8.0 ml. 



anilin blue, C.I. 42780, 1% (I gm./IOO ml. 



water) 2.0 ml. 



pH 4.5 buffer composed of 



O.IM citric acid 1.1 ml. 



0.2M disodium phosphate 0.9 ml. 



distilled water 28.0 ml. 



procedure: 



1. Deparaflinize and hydrate slides to water; remove HgCU. 



2. Stain, start staining at room temperature, and place at once in 

 oven at 60°C: 1 hour. 



3. Wash in running water: 5 minutes. 



4. Dehydrate and clear through 50%, 80% and anhydrous acetone, 

 acetone-xylene (1:1), xylene, 2 changes, and mount. 



results: 



beta cell granules — blue-black 

 acidophil granules — dark red 



