Sih'cr Impregnnlion for Reticuhnn 1(S,') 



4. Bleach in oxalic acid, 5% (5 gm./lOO ml. water); ?) minuies or 

 until clear. 



5. W^ash in running water: 10 minutes. 



6. Wash in distilled water, 3 changes: 5-10 minutes total. 



7. Impregnate in ammoniacal silver carbonate, 50°C: 5 minutes. 



8. Rinse in distilled water, several dips. 



9. Reduce in formalin, 1% (1 ml./99 ml. water), 2 changes: 3 

 minutes total. 



10. Rinse in distilled w^ater, fe^v dips. 



11. Tone in gold chloride imtil sections turn purplish grey. 



12. Rinse in distilled \\ater, few dips. 



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



14. W'^ash in running water: 5-10 minutes. 



15. Dehydrate, clear, and moimt. 



results: 



rcticukim — black 



collagen — reddish purple 



formalin-fixed nuclei — black; cytoplasm — colorless 



BoLiin-fixed nuclei — colorless; cytoplasm — blackish 



Lillie's Method (1946) 



fixation: any good general fixative. 



solutions: 



Ferric chloride: 



ferric chloride, official solution (29.1%) 1.0 ml. 



distilled water 49.0 ml. 



Ammoniacal silver hydroxide: 

 ^ Use acid-clean glassware. Place 1 ml. ammonia (28% reagent) in small 

 flask; add 7-8 ml. silver nitrate 10% (10 gm./lOO ml. water) rapidly. 

 Continue to add siher nitrate drop by drop, shaking between each 

 addition until a faint turbidity remains (approximately 9-10 ml. 

 silver nitrate). Dilute restiltant solution with equal ainount of dis- 

 tilled water. 



procedure: 



1. Deparaffinize and transfer slides to absolute alcohol. 



2. Coat with celloidin; drain; harden in 80% alcohol. 



3. Hydrate slides to water; remove HgCU. 



4. Oxidize in potassium pennanganate, 0.5% (0.5 gm./lOO mk 

 water): 2 minutes. 



