Pictures will be available soon.

TECHNIQUE:        Undecalcified glycol methacrylate embedded bone marrow sections cut at 4 μm.

SOLUTIONS:

Modified Weigert’s Iron Hematoxylin

Solution A

Hematoxylin, C.I. 75290 -------------------------------------------------------        -        2.0 gm

Alcohol, 90% --------------------------------------------------------------------        100.0 ml

Solution B

Ferric chloride, FeCl3 . 6H20, 62% aqueous -----------------------------------                4.0 ml

Distilled water ----------------------------------------------------------------                95.0 ml

Hydrochloric acid, concentrated ----------------------------------------------                1.0 ml

Working Modified Weigert’s Iron Hematoxylin

Equal parts of Solution A and Solution B

Biebrich Scarlet-Acid Fuchsin Solution

Biebrich scarlet, C.I. 26905 ----------------------------------------------------                0.45 gm

Acid fuchsin, C.I. 42685 -------------------------------------------------------                0.05 gm

Acetic acid, glacial --------------------------------------------------------------                0.50 ml

Phosphomolybdic-Phosphotungstic Acid Solution

Phosphomolybdic acid ----------------------------------------------------------                2.5 gm

Phosphotungstic acid -----------------------------------------------------------                2.5 gm

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

Aniline Blue Solution

Aniline blue, C.I. 42755 --------------------------------------------------------                1.0 gm

Distilled water ------------------------------------------------------------------                40.0 ml

Acetic acid, glacial --------------------------------------------------------------                0.8 ml

1% Acetic Acid Solution

Acetic acid, glacial --------------------------------------------------------------                1.0 ml

Distilled water ------------------------------------------------------------------                99.0 ml

1. Place in modified Weigert’s iron hematoxylin for 5 minutes.
2. Wash briefly in running tap water and rinse in two changes of distilled water.
3. Decolorize with 0.5% hydrochloric acid in 70% alcohol for 5 seconds.
4. Wash in running tap water for 30 seconds and rinse in two changes of distilled water.
5. Place in biebrich scarlet-acid fuchsin solution for 30 minutes.
6. Rinse in three changes of distilled water.
7. Place in phosphomolybdic-phosphotungstic acid for 10 minutes.
8. Rinse in two changes of distilled water.
9. Place in aniline blue solution for 7 minutes.
10. Rinse in two changes of distilled water.
11. Place in 1% acetic acid for 1 minute.
12. Rinse in three changes of distilled water and air dry.
13. Dip in xylene and mount with synthetic resin.

Osteoid seams -------------------------------------------------------        red

Mineralized bone ---------------------------------------------------        blue

Nuclei ----------------------------------------------------------------        dark gray

Masson, P.J.:  Some histological methods.  Trichrome stainings and their preliminary technique.  J. Tech. Methods 12:75-90, 1929.

Elbadawi, A.:  Hexachrome modification of Movat’s stain.  Stain Technol. 51:249-253, 1976.