TECHNIQUE:        Undecalcified glycol methacrylate embedded sections cut at 4 μm or paraffin sections cut at 5 μm.

SOLUTIONS:

Buffer Solution, pH 5.2

Ammonium acetate -------------------------------------------------------------                40.0 gm

Ammonium chloride ------------------------------------------------------------                28.0 gm

Distilled water ------------------------------------------------------------------                210.0 ml

50% Hydrochloric acid, (6N) --------------------------------------------------                27.0 ml

       Adjust to pH 5.2 with concentrated hydrochloric acid or 28%

       ammonium hydroxide.  Store in refrigerator at 3-6o C.

*Aluminon Solution

Aluminon -------------------------------------------------------------------------        0.8 gm

Buffer solution, pH 5.2 ---------------------------------------------------------        --        40.0 ml

       Place in a 125 ml flask and place on a hot plate stirrer.  Heat to

            80-85o C just before use.

       *See Step 2 of the Staining Procedure.

1.6M Ammonium Carbonate

Ammonium carbonate ----------------------------------------------------------                15.4 gm

Distilled water to make a total of ------------------------------------------------                100.0 ml

       Store in a refrigerator at 3-6o C.

Differentiating Solution, pH 7.2

Buffer solution, pH 5.2 ---------------------------------------------------------                22.0 ml

1.6M ammonium carbonate ----------------------------------------------------                8.0 ml

Fast Green, Stock Solution

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

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

Acetic acid, glacial --------------------------------------------------------------                0.2 gm

Fast Green, Working Solution

Fast green, stock solution ------------------------------------------------------                10.0 ml

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

1. Place GMA embedded slides on a hot plate at 60-65o C for 15 minutes.  Deparaffinize and hydrate the paraffin sections with distilled water.
2. Heat the aluminon solution in a 125 ml flask to 80-85o C on a hot plate stirrer.
3. Pour the hot aluminon solution in a plastic Coplin jar and place the slides in the solution.
4. Place Coplin jar in a microwave oven and microwave at power level 2 (120 W) for 30 seconds.  Dip the slides up and down and allow them to remain in the hot solution for 7 minutes.
5. Rinse in distilled water.
6. Place in differentiating solution for 5 seconds.
7. Rinse quickly in three changes of distilled water.
8. Counterstain with working fast green solution for 3 minutes.
9. Rinse in three changes of distilled water and air dry.
10. Dip in xylene and mount with synthetic resin.

Aluminum ----------------------------------------------------------        red

Background -------------------------------------------------------                varying shades of green

Lillie, R.D.:  Histopathologic Technic and Practical Histochemistry, 4th ed., New York, McGraw-Hill, 1976, p. 534.

Ohtsuki, T. et al.: A simplified aluminum stain in paraffin sections of bone from hemodialysis patients. Stain Technol. 64: 55-59, 1989.