Mallory-Heidenhain Azan-Gomori’s Modification for Islet Cells

EMS26450

Components

Azocarmine Stain Solution

Aniline-Alcohol 1%

Ferric Ammonium Sulfate 5%

Aniline Blue-Orange Solution

Orange G 0.2% In Alcohol

Fixation

Bouin's or Helly's

Section

Paraffin, 3-5µm

Staining

1. Deparaffinize and hydrate to distilled water as usual.
2. Wash in running water to remove all picric acid.
3. Stain in Azocarmine G 0.1% 45-60 Min. Perform in a covered petri dish at 56°C. Rinse and blot dry carefully. (NOTE: Azocarmine G has been restricted in use as a carcinogen and not recommended, Azocarmine B may possibly be substituted depending upon material to be stained.)
4. Differentiate in the Aniline-Alcohol, 1% according to fixation and species used. Check periodically under a microscope until the beta cells are stained either orangy red with human tissue or red with guinea pig tissue. The beta cells should appear red against the pinkish alpha cells with colorless parenchyma (acinar).
5. Rinse in distilled water.
6. Transfer to Iron Alum Solution for 5 minutes or more- rinse in distilled water.
7. Stain in Aniline Blue-Orange G for 2 to 20 minutes under microscopic control. Rinse briefly and blot carefully.
8. Differentiate in absolute alcohol and dehydrate completely. (NOTE: if the orange colour fades too much in this step, restain the slide in Orange G, 0.2% saturated alcohol, and rewash and redehydrate with clean absolute alcohol.)
9. Clear with xylene, two changes each.
10. Mount.

Stain Results:

(Bouin Fixative)

References

Clark, G, ed.: Staining Procedures 3rd. ed.; Baltimore: Williams and Wilkins Co., c. 1973, p171.
 Gomori. G: "Studies on the cells of the Pancreatic Islets." Anat Rec. Rec. 74: 439 (1939).