Sample type required: Deparaffinized and rehydrated tissue section (10 μ) on positively (+) charged slides
Preferred fixative: 10% neutral buffered formalin (NBF)
Control: Normal cerebral cortex, spinal cord, or medulla
|Luxol fast blue (LFB) solution||Overnight in 58° C oven|
- Be sure to cap/seal the coplin jar to prevent evaporation of the solution.
|95% alcohol||1 change|
- Rinse to remove excess stain.
|Distilled water||1 change|
|*Lithium carbonate solution||10 seconds|
- Use for differentiation.
- Be careful not to over-differentiate.
|*70% alcohol||2 changes|
- Rinse to further differentiate and remove lithium carbonate solution.
|*Distilled water||3 changes|
|Repeat the previous 3 steps denoted by the (*)||Until there is a sharp contrast between the blue of the white-matter and the colorless gray-matter.|
- Check microscopically for differentiation.
- Be careful not to over differentiate.
Post staining procedure: Tissue section should be dehydrated with 95% and absolute alcohols. Follow with 2 changes of xylene and then coverslip.
- Myelin - Blue
- Background - Colorless
Image courtesy of Wikimedia Commons.