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Silver Stain (Modified Steiner)

Reagent Preparation

Prepare working Hydroquinone solution by dissolving one hydroquinone tablet in 25 mLs of distilled water. Mix well. Prepare Reducing solution by mixing 10 ml Gum Mastic solution, 25 mL working Hydroquinone, and 5 mL Absolute alcohol. Filter solution. Discard Uranyl Nitrate, Silver Nitrate, and Gum Mastic solutions if turbidity develops.

Procedure

  1. Preheat 40 mL of Uranyl Nitrate solution and 40 mL Silver Nitrate solution to 60 °C.
  2. Deparaffinize tissue sections and hydrate to deionized water.
  3. Place in Uranyl Nitrate solution in 60 °C water bath for 15 minutes.
  4. Rinse in 3 changes of deionized water.
  5. Place in Silver Nitrate solution in 60 °C water bath for 1.5 hours.
  6. Rinse in 3 changes of deionized water.
  7. Rinse in 2 changes each of 95% and absolute alcohol.
  8. Place in Gum Mastic solution for 5 minutes.
  9. Remove slides from Gum Mastic solution and allow sections to air dry.
  10. Prewarm Reducing solution to 45 °C in water bath.
  11. Place in Reducing solution and incubate for approximately 25 minutes.
  12. Remove coplin jar from water bath and allow slides to stand in Reducing solution at room temperature until desired stain intensity is achieved, 1-2 minutes is optimal. Longer staining is optional, if desired.
  13. Dehydrate in alcohol, clear in xylene and mount.

Results

Spirochete – Brownish black
Helicobacter – Brownish black
Background – Bright yellow to light brown

Silver Stain (Modified Steiner)

Reagents

     
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