Woelck’s Method for Myelin Sheath

EMS26684

Components:

Ferric Ammonium Sulphate 2.5% Aqueous

Alcoholic Haematoxylin Stock 10%

Lithium Carbonate Saturated Solution

Fixation:

Formalin 10% Neutral Buffered

Sections:

Paraffin @ 15µm

Staining Procedures:

1. Deparaffinise and hydrate to distilled water.
2. Mordant in Ferric Ammonium Sulphate, 2.5% overnight. Rinse in two changes of distilled water.
3. Working Haematoxylin solution for 2 hours. To prepare working solution mix the following ingredients in order listed just before use:
   45.0ml Distilled water
   10.0ml Haematoxylin Stock – filtered
   7.0ml Lithium Carbonate, Saturated – filtered
    45.0ml Distilled water
4. Rinse in two changes of distilled water.
5. Differentiate in 80% Alcohol until background is clear and myelin stands out sharply.
6. Dehydrate with 95% Alcohol, absolute alcohol, and clear in Xylene two changes each. Mount.

Stain Results:  

References:

AFIP, Manual of Histological Staining Methods, 3rd ed., Ed. L. Luna: New York: McGraw-Hill Publications. C. 1968, p. 206.