Hale’s Dialyzed Iron Technique

Reagents and Preparation

• Dialyzed Iron

1. 29% aqueous FeCl₃: 22 ml
2. Distilled water: 125 ml

Pour the FeCl₃ solution into the boiling distilled water and stir well. The solution will turn dark red when it is allowed to cool. Ensure that the distilled water is boiling when the FeCl₃ is added. Store the solution at 4^o C.

• 2 molar acetic acid
• 2% aqueous potassium ferrocyanide
• 2% HCI
• Schiff’s Reagent

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) Treat all sections with an equal amount of dialyzed iron and acetic acid for 10 minutes.

6) Wash several times in distilled water.

7) Treat with filtered Perl’s solution for 30 minutes.

8) Wash again with distilled water.

9) Either counterstain with 0.5% aqueous neutral red for 5 minutes (preferably 1%) or carry out PAS technique (no hematoxylin).

10) Wash in water.

11) Rinse with absolute alcohol.

12) Clear & mount.

Result

Image Source: VitroVivo Biotech