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BODIAN'S METHOD FOR NERVE FIBERS AND NERVE ENDINGS
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FIXATION: 10% buffered neutral formalin.
TECHNIQUE: Paraffin sections at 6-8 µm.
SOLUTIONS:
1% Protargol Solution
Protargol, certified --------------------------------------------------------------- 0.4 gm
Distilled water -------------------------------------------------------------------- 40.0 ml
Sprinkle the protargol on the surface of the water and allow it to
remain undisturbed until it dissolves.
Reducing Solution
Hydroquinone -------------------------------------------------------------------- 0.4 gm
Formalin, 37-40% --------------------------------------------------------------- 2.0 ml
Distilled water -------------------------------------------------------------------- 40.0 ml
0.2% Gold Chloride Solution
Gold chloride --------------------------------------------------------------------- 0.2 gm
Distilled water -------------------------------------------------------------------- 100.0 ml
2% Oxalic Acid Solution
Oxalic acid ------------------------------------------------------------------------ 2.0 gm
Distilled water -------------------------------------------------------------------- 100.0 ml
2% Sodium Thiosulfate Solution
Sodium thiosulfate --------------------------------------------------------------- 2.0 gm
Distilled water -------------------------------------------------------------------- 100.0 ml
1. Deparaffinize and hydrate to distilled water.
2. Place slides in protargol solution. Add 4 gm of clean copper shot to 40 ml of solution. Allow to remain in a 55-60° C oven overnight.
3. Rinse in distilled water, three changes.
4. Reducing solution for 10 minutes.
5. Rinse in distilled water, three changes.
6. Tone in gold chloride solution for 10 minutes.
7. Rinse in distilled water, three changes.
8. Develop in oxalic acid solution for 4-6 minutes. Check with a microscope until background is gray and nerve fibers appear clearly.
9. Rinse in distilled water, three changes.
10. Sodium thiosulfate solution for 5 minutes.
11. Rinse in distilled water.
12. Dehydrate in graded alcohols.
13. Clear in xylene, three or four changes.
14. Mount with synthetic resin.
Nerve fibers --------------------------------------------------------- black
Nuclei ---------------------------------------------------------------- black
NOTE:
If desired, counterstains may be used. See Manual of Histologic Staining Method of the Armed Forces Institute of Pathology, 3rd ed., New York, McGraw-Hill, 1968, pp. 195-196.
The glassware should be thoroughly cleaned and rinse with double distilled water.
Luna, L.G.: Further Studies of Bodians Technique. Amer. J. Med. Techn. 30:355-363, 1964.
Mallory, F.B.: Pathological Technique, New York, Hafner, 1968, pp. 228-229.
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