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Nissl body, also known as Nissl substance, is a large granular body located in cytoplasm of neurons. The Nissl body major contains ribosome and rough endoplasmic reticulum, is the site of protein synthesis. Distribution of Nissl body is a marker demonstrating neuron injury or not. This staining is commonly used for identifying the basic neuronal structure. This staining method is based on toluidine blue staining.
1) Paraffin section deparaffinate: put sections into xylene Ⅰ 20min- xylene Ⅱ 20min- anhydrous ethanol Ⅰ 5min- anhydrous ethanol Ⅱ 5min-75% alcohol 5min,then washing in tap water.
2) Staining in Nissl stain solution for 10 min.
3) Differentiate in 95% alcohol (proper differentiate extent for brain tissue is that Nissl body shows deep blue, while background is light blue or colorless ), wash in tap water.
4) Dry in the air and mount with mounting medium .
Nissl bodies are shown in deep blue, nucleus are light blue and background is colorless.