This product is used to stain myelin sheath by luxol fast blue. Myelin sheath is composed of sphingomyelin, with lipoids and proteins as the major component. It is the insulating covering that surrounds an axon with multiple spiral layers of myelin, discontinuous at the nodes of Ranvier. Due to this characteristic, myelin sheath decreases capacitance and increases the speed at which a nerve impulse can travel along an axon, plays important role in protecting axon in central and peripheral nervous system. Luxol fast blue stain will show normal myelin sheath.
1. Deparaffinize and rehydrate sections to distilled water.
2. Myelin sheath stain: preheat luxol fast blue (LFB) solution in 60℃ oven for 30 min. Immerse slides in preheated stain solution for 3-4 h, cooling for 10-15 min. Wash in running tap water.
3. Differentiate with alternative using of 70% alcohol and lithium carbonate solution. Stop differentiate by wash in tap water. Manage differentiate extent by microscopy observation.
4. Dehydrate and mount: dehydrate in absolute alcohol for 3 times, clear with xylene for 2 times, and then mount with neutral gum.
5. Microscopy detection and analysis.
Myelin sheath is blue, and the other components and background is colorless.
1. Control the washing time after staining, washing too long will fade the blue.
2. Manage the differentiate process, over-differentiate may fade the blue of myelin sheath and inadequate differentiate results in poor clearness.
3. In case of over-differentiate, re-stain slides in LFB solution.