Myelin Sheath is a membrane wrapping around the axons of nerve cells, i.e., it is composed of myelinating cells and cell membranes, forming a multi-layered lipid bilayer structure by the plasma membrane of neurolemmocytes spirally winding along the axis of axons. The myelin sheath has nodes of Ranvier, which enable the saltatory conduction of nerve impulses. Myelin staining is of certain significance in pathological diagnosis. Pathological changes of the myelin sheath are divided into early, intermediate and late stages. In the early stage, the staining is relatively deep; in the intermediate stage of lesions, myelin degeneration forms lipid droplets, which can be displayed by lipid staining; in the late stage, the myelin sheath completely degenerates and is cleared by phagocytes, so there will be no positive result of myelin sheath.
Many diseases can cause changes in the myelin sheath. Modified Page Myelin Staining Solution, also known as Gallocyanin Myelin Staining Solution, can display the integrity, degeneration, necrosis degree and repair status of the myelin sheath under pathological conditions, which is of significance for the pathological diagnosis and research of nerve tissues. The myelin sheath appears blue, while demyelinated fibers are unstained. This reagent is for research use only and not intended for clinical diagnosis or other purposes.
Materials to Be Prepared by User
Distilled water, gradient ethanol, xylene or dewaxing clearing solution, neutral mounting medium
Procedure (for reference only)
1. Fix the tissue in Page Formaldehyde Fixative for more than 3 days.
2. Perform routine dehydration and embedding, slice into sections with a thickness of 5 μm, then dewax the sections to distilled water using xylene or dewaxing clearing solution.
3. Stain the sections with Modified Page Staining Solution for 15-25 minutes.
4. Pour off the staining solution and rinse the sections with running water for 1 minute.
5. Take an appropriate amount of Page Differentiation Solution, prepare Page Differentiation Working Solution at a ratio of Page Differentiation Solution: distilled water=1:1; differentiate the sections directly with the working solution for 20-180 seconds. The differentiation process should be observed under a microscope to control the differentiation degree.
6. Rinse the sections with tap water for 10 minutes.
7. Counterstain the sections with Page Fuchsine Staining Solution for 3-6 seconds (see Precaution 3), then rinse briefly with tap water.
8. Perform routine dehydration with gradient ethanol, clear the sections with xylene or dewaxing clearing solution, and mount with neutral mounting medium.
Staining Results
Myelin sheath, cell nuclei: Dark blue
Red blood cells: Green
Cytoplasm, collagen fibers, muscle fibers: Fuchsine
Precautions
1. Differentiation is a critical step, and the differentiation time should be strictly controlled. The differentiation degree can be observed under a microscope. You can differentiate for a few seconds first, rinse with water and observe under the microscope until the collagen fibers and muscle fibers are nearly colorless or light gray, and the myelin sheath shows clear blue.
2. If the differentiation process is difficult to control, the Page Differentiation Solution can be diluted 4-5 times with distilled water before differentiation.
3. The sections should not be too thick, and the thickness should be controlled within 5-6 μm; otherwise, section detachment or over-staining may occur.
4. New Page Fuchsine Staining Solution requires a relatively short staining time, while the staining ability of the stored solution for a long time will decrease, and it can be slightly warmed before staining for several minutes.
5. For your safety and health, please wear a lab coat and disposable gloves during operation.
6. Please use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.
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