Silver Impregnation for Reticiilum 187 



10. Wash in distilled water: 2-3 minutes. 



11. Tone in gold chloride (10 ml. 1% stock/80 ml. water) until yel- 

 low color turns pmplish grey. 



12. Rinse briefly in distilled water. 



13. Fix in sodium thiosulfate, 5% (5 gm./lOO ml. water): 3 minutes. 



14. Wash in running water: 5 minutes; rinse in 70% alcohol. 



15. Stain in orcein solution, 37 °C: 15 minutes, or at room temper- 

 ature: 1 hour. 



16. Rinse in 70% alcohol, followed by distilled water. 



17. Treat with phosphomolybdic acid, 5% (5 gm./lOO ml. water): 

 10-15 minutes. 



18. Briefly rinse in distilled water. 



19. Stain in aniline blue solution: 2-3 minutes. 



20. Rinse off some of blue in distilled water, and transfer to acidu- 

 lated water (1 ml. glacial acetic acid/ 100 ml. water): 5 minutes or 

 longer. 



21. Dehydrate, clear, and mount. 



To modify with the addition of orange G, proceed after step 19 with 

 a rinse in distilled water and continue as follows: 



20. Stain in orange G (C.I. 16230) 2% (2 gm./lOO ml. water): 1-2 

 minutes. Extended time didls aniline blue. 



21. Rinse in distilled water and treat with acididated water. 



22. Dehydrate, clear, and mount. 



results: 



reticulum — black 



elastin — red 



collagen— blue 



colloid — brown or grey 



nuclei — brown-black 



erythrocytes — orange 



smooth muscle — -beige 



striated muscle — grey-blue or mauve 



epithelium — light grey-brown 



comments: 



The orange G can be added to the aniline blue solution but separate 

 solutions offer better individual control of each color intensity. If 

 sections tend to loosen, follow step 14 by dehydrating slides to abso- 

 lute alcohol, apply a coat of celloidin, harden it in 70% alcohol and 

 proceed to step 15. 



