Sample type required: Deparaffinized and rehydrated tissue section (10 μm) on positively (+) charged slides
Preferred fixative: 10% neutral buffered formalin (NBF)
Control: Normal cerebral cortex, spinal cord, or medulla
| Reagent | Time | Technical Notes |
| Luxol fast blue (LFB) solution | Overnight in a 58° C oven | - Be sure to cap/seal the coplin jar to prevent evaporation of the solution.
|
| 95% alcohol | 1 change (at least 4 dips) | - Rinse to remove excess stain.
|
| Distilled water | 1 change | |
| *Lithium carbonate solution | 10 seconds (at least 4 dips) | - Use for differentiation.
- Be careful not to over-differentiate.
|
| *70% alcohol | 2 changes | - Rinse to further differentiate and remove the 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: The tissue section should be dehydrated through graded alcohols, beginning with 95%. Followed by 2 changes of xylene and then a coverslip.
Expected results:
- Myelin - Blue
- Background - Colorless
