Reagents and Preparation
• Solution A: 5 grams of hematoxylin dissolve in 100 ml of absolute alcohol on a hot plate, cool, and filter.
• Solution B: 10 grams of ferric chloride dissolve in 100 ml of distilled water.
• Solution C: 1 gram of iodine & 2 grams KI dissolve in 100 ml distilled water .
• Solution D or working solution
20 ml of solution A + 8 ml of solution B + 8 ml of solution C.
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) Expose to elastic stain for 15–30 minutes.
6) Rinse with distilled water.
7) Differentiate in 2% ferric chloride for a few minutes (till elastic tissue is stained black in gray).
8) Rinse with distilled water.
9) Counter stain with Van Gieson’s stain for 1-2 minutes.
10) Blot dry, clear in xylene, and mount.
Result
Elastic fiber: black
Muscle: yellow
Collagen: red
Image Source: X