Mast cells are derived from undifferentiated mesenchymal cells and are found in tissues such as perivascular submucosa, loose connective tissues, tumor tissues, and allergic diseases, and are easily colored by heterochromatic dyes. Source: Immunohistochemistry Experimental Techniques and Applications

Operation method

basic program

Principle

Mast cells have a large morphology (20-30 μm), a small nucleus, and a cytoplasm filled with numerous round basophilic granules. Since the granules contain heparin and other components of polysulfated lipids, which also belong to the class of sulfated mucopolysaccharides, they are able to be colored by heterochromatic dyes, and are commonly used for toluidine blue staining.

Materials and Instruments

Paraffin Tissue Sections
Xylene Anhydrous ethanol Neutral gum Distilled water Glacial acetic acid Toluidine blue
Dropper

Move

Toluidine blue staining solution: Toluidine blue 0.5 g, distilled water 100 ml.

Glacial acetic acid differentiation solution: 0.5 ml of glacial acetic acid, 100 ml of distilled water.

1. 4 μm paraffin tissue section, dewaxed to water. 2.

2. Toluidine blue staining solution for 10 min, wash with distilled water twice. 3.

3. differentiate with glacial acetic acid for about 30 s, until the particles are clear (microscope control). 4.

4. Wash with distilled water for 2 times and dehydrate quickly with anhydrous ethanol. 5.

5. Absorb on filter paper and seal with xylene clear and neutral gum.

Caveat

1. Toluidine blue liquid coloring power is strong, according to different tissues, can be appropriate to shorten the staining time.

2. After the action of glacial acetic acid differentiation solution, observe with high magnification of microscope if necessary.

Common Problems

RESULTS: Mast cell granules were purplish red and nuclei were blue.

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