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Jalowy’s Impregnation for Reticulin

Jalowy's Impregnation

for Reticulin

8
steps
4
materials

Materials

  • Silver nitrate, 10% aqu.
  • Strong ammonium hydroxide (s.g. 0.88).
  • Ammonium hydroxide, 1% aqu.
  • Formalin, 10% aqu.

Preparation of Jalowy’s Ammoniacal Silver

  1. Place 20 mL of 10% silver nitrate in a flask and add 1 mL of 40% sodium hydroxide.
  2. Mix well, then filter. Wash the precipitate well with distilled water several times and decant.
  3. Add 20 mL distilled water to the precipitate and suspend.
  4. Add strong ammonium hydroxide by drops until the precipitate is just dissolved.
  5. Dilute to 100 mL with distilled water.

Tissue Sample

5 µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory. A section adhesive is recommended.

Protocol

  1. Bring sections to water via xylene and ethanol.
  2. Rinse with distilled water.
  3. Place in prewarmed ammoniacal silver solution for 5-30 minutes at 30°C.
  4. Rinse with distilled water.
  5. Rinse with 1% ammonium hydroxide.
  6. Place in 10% formalin for 2-10 minutes.
  7. Rinse well with tap water.
  8. Dehydrate with ethanol, clear with xylene, and mount with a resinous medium.

Expected Results

  • Reticulin fibres  –  black
  • Background  –  brown

Notes

  • This method was recommended for collagen and reticulin fibres on formalin fixed skin.
  • Ensure that the ammonium hydroxide and sodium hydroxide are fresh and full strength. Keep the ammonium hydroxide well stoppered when not in use. Pour sufficient for use into a beaker, then immediately restopper the stock container. Do not return unused ammonium hydroxide to the stock bottle.
  • The instructions for this method do not specify to tone, fix, or counterstain. If fading or a lack of contrast make it desirable, toning with 0.1% yellow gold chloride until satisfactory, and/or fixing with 5% sodium thiosulphate, and/or counterstaining with 1% aqueous neutral red may be used.

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.

References

  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.