Nissl staining solution (methylene blue method) instruction manual

Nissl staining solution (methylene blue method)

Product introduction:

The neuronal cell body includes a large cell nucleus with a wrinkled nuclear membrane, sparse chromatin, and a distinct nucleolus. The cytoplasm in the cell body is a Nissl particle, an basophilic particle capable of representing a rough endoplasmic reticulum and producing a specific speckled basophilic expression in many neurons. Nissl particles can be marked with many dyes such as neutral red, methylene blue, toluidine blue and methyl violet. The variation of staining, pH, and time of differentiation allow some stains to highlight only Nissl's material, as well as the nucleus and glia of neurons. Nissl body or Nissl body is a triangular or elliptical small block material distributed in the cytoplasm of the nerve. It can be dyed by basic dyes such as thiopurine, methylene blue, toluidine blue and tar violet. Dyed in purple blue. Nissl bodies are contained in various nerve cells, but their shapes, numbers, and locations are often different. Nissl bodies are also present in dendrites, but not in the axons and inclusions. Nissl's body changes due to changes in physiological state. Nissl is an important part of synthetic protein synthesis in neurons. When neurons are stimulated, the Nissl body in the body will be significantly reduced.

Nissl Stain (Nissl Stain, methylene blue method) has the advantages of simple operation, stable dyeing and wide application range. It can be used for dyeing Nissl substances and neurons of paraffin tissue sections, and the presence and disappearance of Nissl. It is an important indicator of whether nerve cells are damaged. When encephalitis, cerebral ischemia, axon reaction, etc. occur, the Nissl body will dissolve or even disappear.

product composition:

Bring your own materials:

1, 20% formaldehyde solution,

2, microscope

3, distilled water

Operation steps (for reference only):

1. Fresh tissue was fixed in 20% formalin and routinely dehydrated.
2. The sections were 5 μm thick and were conventionally dewaxed to water.
3. Methylene Blue Stain is dyed for 10min.
4. Differentiation into Nissl Differentiation for 1~3 min, observed under the microscope until the Nissl body is clear.
5. The sections were processed in Ammonium Molybdate Solution for 5 min.
6. Rinse with distilled water.
7. Conventional dehydration is transparent and sealed by neutral gum.

Dyeing results:

Precautions:

1. Nissl bodies are easily dissolved after being isolated, so the tissue should be fixed immediately after removal, otherwise it is difficult to color.
2. Tissue fixation plays a very important role, and ethanol, Carnoy fixative or neutral formalin solution can be used for fixation.
3. The staining kit has a better effect on the Nissl staining of paraffin tissue sections.
4. Paraffin sections were 7–10 μm or 25 μm thick (the assessment of cortical neuron density was performed with 25 μm thick sections).
5. The stained specimen must be stored away from light, otherwise it will fade easily.
6. For your safety and health, wear a lab coat and disposable gloves.

Validity: Valid for 6 months.

If you encounter any technical problems in the experiment, please consult the company's technical staff in time.