Reagents
- Potassium permanganate: 1%
- Sodium or potassium metabisulfate: 3%
- Iron Alum: 2%
- Formalin: 10%
- Gold Chloride: 0.2%
To make a silver solution, the following reagents are required:
- Silver nitrate: 10%
- Potassium hydroxide: 3%
- Liquor ammonia: Concentrated
- Distilled water
Make the titrated silver solution in a test tube freshly before each & every stain (12 drops of AgNO3 + 3 drops of KOH + add drop by drop NH3 till the precipitate dissolves completely & gives a colorless solution + again add 6 drops of AgNO3 + add an equal volume of distilled water to the solution).
Staining Procedure
1) Deparaffinize the slide by heating in hot incubation.
2) To remove paraffin, the slide has to be dipped in xylene for 1-2 minutes. Repeat the process four times.
3) Keep the slide in alcohol for 1-2 minutes to remove xylene. Repeat the process two times.
4) Wash the slide in running water.
5) Cover the slide with 1% KMnO4 and keep it for 2 minutes.
6) Drain off the excess KMnO4 and wash in distilled water.
7) Add 3% sodium metabisulfate and wait for 2–3 minutes.
8) Drain off the excess and wash in distilled water.
9) Cover the slide with 2% iron alum and wait for another 5 minutes.
10) Drain off the excess and rinse with distilled water.
11) Titrated solution is given to the slide for 5 minutes.
12) Excess is drained off and washed in distilled water.
13) Keep the slide in 2% formalin for 2 minutes.
14) Rinse in distilled water.
15) Add 0.2% Gold Chloride and wait for 5-10 minutes.
16) Wash in running water.
17) Dry up xylene and mount.
Results
Reticulin fibres appear black.
Image source: ResearchGate