Perls Iron Stain

From:Steven Slap

Hi HistoNetters

Neelima Shah asks for a Perls Iron Stain protocol.  There is a good one in
Kok & Boon's Microwave Cookbook for Pathologists:

Procedure 14.15 The Perls Iron Stain

the Perls Iron stain is a metallic-staining method widely used to
demonstrate ferric iron, for instance in bone marrow.  The conventional
method takes 20 min, in the microwave method it can take less than 15 s.


To stain ferric iron.

Solutions needed:

 Working solution:
1 ml 2% Potassium ferrocyanide
1 ml 2% Hydrochloric acid
Prepare fresh before use
 Nuclear fast red solution:
0.1 g Nuclear fast red
100 ml 5% aluminum-sulfate solution
Boil for 5 min.  Add grain of thymol as preservative.

Staining procedure:

1. Deparaffinize and hydrate to distilled water.
2. Place slide in plastic staining jar containing 16 ml perls working
solution and microwave at 450 W for 15 s (for 50 ml Perls working solution
the microwave-exposure time is 50 s).
3. Rinse well in distilled water.
4. Counterstain in 16 ml nuclear fast red at 450 W (60C if using a
temperature probe) for 15 s.
5. Wash well in running water.
6. Dehydrate in 96% alcohol, absolute alcohol, clear in xylene two changes
each, and mount.


Use chemically clean glassware and control slide.


ferric iron: bright blue
nuclei: red

Perls, M (1867), Nachweis von Eisonoxyde in Pigmenten. Virchow Arch Path
Anat 39:42-28.

best regards,
Steven Slap

Marketing Manager/Microwave Product Specialist
Hacker Instruments & Industries, Inc.

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