Reticulum Stain Kit (Brenn)
Preparation of Reagents Prior to Beginning:
1. Prepare working Ammoniacal Silver Solution using chemically cleaned glassware in a chemical fume hood as follows: To 17ml of Distilled Water add one (10ml) vial of Silver Nitrate Solution (10%) and mix completely. Add 1ml of Concentrated Ammonium Hydroxide (Not Included in Kit) while continuously mixing. The mixture will initially turn brown and then become clear. Add 10ml of Sodium Hydroxide Solution (3%) and mix completely. If the solution does not remain colorless, add Concentrated Ammonium Hydroxide drop by drop until no precipitate remains. Add Distilled Water to a total volume of 60ml and mix completely. Solution is now ready for use.
Note: Use extreme care in preparation and use of Ammoniacal Silver Solution. Store Ammoniacal Silver Solution in a refrigerator to avoid the formation of explosive compounds. If Ammoniacal Silver Solution is exposed to sunlight, it will explode. Dispose of waste observing all local, state and federal laws.
1. Deparaffinize sections if necessary and hydrate to distilled water.
2. Place slide in Potassium Permanganate Solution (1%) for 5-10 minutes.
3. Rinse in 3 changes of distilled water.
4. Differentiate in Potassium Metabisulfite Solution (3%) until section is transparent.
5. Rinse in 3 changes of distilled water.
6. Apply Ferric Ammonium Sulfate Solution (3%) for 10 minutes.
7. Rinse in 2 quick changes of distilled water.
8. Apply working Ammoniacal Silver Solution for 2-3 minutes.
9. Rinse in 3 changes of distilled water.
10. Place slide in Formalin Solution (20%) for 1 minute.
11. Rinse in 3 changes of distilled water.
12. Apply Gold Chloride Solution (0.1%) for 3-5 minutes.
13. Rinse in 2 changes of distilled water.
14. Apply Sodium Thiosulfate Solution (5%) for 1-2 minutes to remove unreduced silver.
15. Rinse in tap water for 2 minutes.
16. Counterstain using Nuclear Fast Red Solution for 2-5 minutes.
17. Rinse in tap water followed by distilled water.
18. Dehydrate through 3 changes of Absolute Alcohol.
19. Clear, and mount in synthetic resin.
Hover over each placard to see a description of its meaningUN1198 Formaldehyde solutions, flammable, 3, III