Jalowy's Impregnation
for Reticulin
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
- Place 20 mL of 10% silver nitrate in a flask and add 1 mL of 40% sodium hydroxide.
- Mix well, then filter. Wash the precipitate well with distilled water several times and decant.
- Add 20 mL distilled water to the precipitate and suspend.
- Add strong ammonium hydroxide by drops until the precipitate is just dissolved.
- 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
- Bring sections to water via xylene and ethanol.
- Rinse with distilled water.
- Place in prewarmed ammoniacal silver solution for 5-30 minutes at 30°C.
- Rinse with distilled water.
- Rinse with 1% ammonium hydroxide.
- Place in 10% formalin for 2-10 minutes.
- Rinse well with tap water.
- 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
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.