Silver Impregnation for Reticulum 170 



Silver Impregnation for Reticulum 



Bielschowsky-Foot Method (mallory, 1938) 

 fixation: 10% formalin or Zenker-formol. 



solutions: 



Ammoniacal silver solution: 



In acid-clean glassware place 20 ml. of 10% silver nitrate (10 gm./lOO 

 ml. water); add 20 drops of 40% sodium hydroxide (40 gm./lOO ml. 

 water). Dissolve precipitate ^vhich forms with ammonia, drop by drop, 

 with shaking until the precipitate is almost dissolved. Filter out re- 

 maining granules and make up to 80 ml. with distilled water. Make 

 up fresh each time. 



Gold chloride: 



gold chloride, 1% stock solution (1 gm./lOO ml. 



water) 1.0 ml. 



distilled water 99.0 ml. 



procedure: 



1. Deparaffinize and hydrate slides to water: remove HgCU. 



2. Rinse in distilled water. 



3. Oxidize in potassium permanganate, 0.25% (0.25 gm./lOO ml. 

 water): 5 minutes. 



4. Wash in running water: 3 minutes. 



5. Bleach in oxalic acid, 5% (5 gTn./lOO ml. water) till clear. 



6. Wash in rimning water: 5 minutes. Rinse in distilled water. 



7. Treat in silver nitrate, 2% (2 gm./lOO ml. water): 48 hours, in 

 subdued light, not dark. 



8. Rinse in distilled water. 



9. Impregnate with ammoniacal silver: 30 minutes. 



10. Rinse very quickly in distilled water. 



11. Reduce in formalin 5% (5 ml./95 ml. water): 30 minutes. 



12. Rinse in tap water. 



13. Tone in gold chloride until fibers take on blackish grey color. 



14. Rinse in distilled water. 



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



16. Wash in riuining water: 5-10 miniues. 



17. Cotmterstain if desired. 



18. Dehydrate, clear, and mount. 



