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FIXATION:        10% buffered neutral formalin.

TECHNIQUE:        Paraffin sections cut at 5 μm.

SOLUTIONS:

0.5% Periodic Acid Solution

Periodic acid ---------------------------------------------------------------------        0.5 gm

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

Modified Lillie’s Schiff Solution

(see PAS method)

Weigert’s Iron Hematoxylin (Modified)

(see Gomori’s One Step Trichrome method)

0.5% Acid Alcohol Solution

Alcohol, 70% -------------------------------------------------------------------                1000.0 ml

Hydrochloric acid, concentrated -------------------------------------------------        5.0 ml

1% Orange G Solution

Orange G, C.I. 16230 -----------------------------------------------------------        1.0 gm

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

5% Phosphotungstic Acid Solution

Phosphotungstic acid -----------------------------------------------------------                5.0 gm

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

• Use control slide.

1. Deparaffinize and hydrate to distilled water.
2. Oxidize in 0.5% periodic acid for 10 minutes.
3. Rinse in four changes of distilled water.
4. Lillie’s Schiff solution for 15 minutes.
5. Rinse in three changes of distilled water.
6. Place in 0.3% sodium borate for 15 seconds.
7. Rinse in four changes of distilled water.
8. Modified Weigert’s iron hematoxylin for 1 minute.
9. Rinse in two changes of distilled water.
10. Acid alcohol solution for 10 seconds.
11. Wash in running water and rinse in distilled water.
12. Orange G solution for 1 ½ minutes.
13. Place in 5% phosphotungstic acid for 30 seconds.
14. Wash briefly in running water and rinse in two changes of distilled water.
15. Dehydrate in graded alcohols.
16. Clear in three or four changes of xylene.
17. Mount with synthetic resin.

Beta granules -------------------------------------------------------        red

Gamma granules ---------------------------------------------------        purple

Acidophils ----------------------------------------------------------                yellow-brown

Nuclei ---------------------------------------------------------------        black

The glandular cells of the adenohypophysis may be broadly classified as chromophilic or chromophobic based on their affinity (or lack thereof) of certain dyes.  Chromophilic cells may be subdivided into acidophils or basophils based on the reactions of their specific cell granules with acid or basic dyes.  The chromophobe cells are believed to represent either precursor cells for the chromophils, or chromophils in various stages of degranulation.  The immunocytochemical procedure is the most specific method for demonstrating the various types of cells found in the pituitary.

Wilson, W.D. and Ezrin, E.:  Three types of chromophil cells of the adenohypophysis, demonstrated by a modification of the periodic acid –Schiff technique.  Amer. J. Path. 30:891-899, 1954.