Reticulum Stain

Reagent Preparation

Ammoniacal Silver Nitrate Solution

  1. Pipet 5 mL Silver Nitrate solution in an Erlenmeyer flask.
  2. While swirling flask, slowly add concentrated ammonium hydroxide, drop by drop, until precipitate it formed is completely dissolved. Do not add excess.
  3. Add 5 mL Sodium Hydroxide solution to flask. Black precipitate will form. Continuously swirl the flask and add concentrated ammonium hydroxide, drop by drop until precipitate dissolves. Do not dissolve until completely clear. Faint cloudiness is desired.
  4. Dilute to 50 mL with deionized water. Filter, use once and then discard.

Procedure

  1. Deparaffinize tissue sections and hydrate to deionized water.
  2. Oxidize in Potassium Permanganate solution for 5 minutes.
  3. Rinse in tap water for 2 minutes.
  4. Bleach in Oxalic Acid solution for 2 minutes or until section are colorless.
  5. Wash in tap water for 2 minutes.
  6. Sensitize in Ferric Ammonium Sulfate solution for 15 minutes.
  7. Rinse in several changes of deionized water.
  8. Impregnate in Ammoniacal Silver Nitrate solution for 2 minutes.
  9. Rinse well in deionized water.
  10. Reduce in 10% Formalin solution for 2 minutes.
  11. Wash in tap water for 3 minutes.
  12. Tone in Gold Chloride solution for 10 minutes.
  13. Rinse in deionized water.
  14. Place in Sodium Thiosulfate solution for 1 minute. Rinse in distilled water for 2 minutes.
  15. Counterstain in a Nuclear Fast Red solution for 3-5 minutes or an Eosin Y solution for 1-2 minutes. Wash in water.
  16. Dehydrate in alcohol, clear in xylene and mount.

Results

Reticulum – Black
Background – Pink to Rose (If counterstained with Nuclear Fast Red)

Reticulum Stain

Reagents

     
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