FIXATION:        10% buffered neutral formalin.

TECHNIQUE:        Paraffin sections cut at 5 μm.

SOLUTIONS:

Lugol’s Iodine Solution

(See Verhoeff’s Elastic Method)

2% Sodium Thiosulfate Solution

Sodium thiosulfate --------------------------------------------------------------                2.0 gm

Distilled water ------------------------------------------------------------------                100.0 ml

Aldehyde Fuchsin Solution

(See Aldehyde Fuchsin Method for HbsAg)

0.2% Fast Green Solution (Stock)

Fast green FCF, C.I. 42053 ----------------------------------------------------                0.2 gm

Distilled water ------------------------------------------------------------------                100.0 ml

Acetic acid, glacial --------------------------------------------------------------                0.2 ml

Fast Green Solution (Working)

Fast green (stock) ---------------------------------------------------------------        10.0 ml

Distilled water -------------------------------------------------------------------        35.0 ml

• Use control slide.

1. Deparaffinize and hydrate to distilled water.  Omit Steps 2-5 if the aldehyde fuchsin is fresh (5-10 days old).
2. Lugol’s iodine solution for 5 minutes.
3. Rinse in two changes of distilled water.
4. Place in 2% sodium thiosulfate for 1 minute.
5. Wash in running tap water and rinse in two changes of distilled water.
6. Place in 70% alcohol for 30 seconds.
7. Aldehyde fuchsin in a covered Coplin jar for 30 to 60 minutes.  Longer staining times may be necessary if the solution is old.
8. Rinse in four changes of distilled water.
9. Fast green working solution for 1 minutes.
10. Rinse in three changes of distilled water.
11. Dehydrate in graded alcohols.
12. Clear in three or four changes of xylene.
13. Mount with synthetic resin.

Beta cells ------------------------------------------------------------        purple to violet

Elastic tissue --------------------------------------------------------        purple to violet

Background ---------------------------------------------------------        green

See COMMENTS under Aldehyde Fuchsin Method for Hepatitis B antigen.

Gomori, G.:  Aldehyde fuchsin:  A new stain for elastic tissue.  Amer. J. Clin. Path. 20:665-666, 1950.