This experimental method was obtained from the official website of Chongqing Medical University

Operation method

Morphological observation experiments on mouse mast cells

Principle

The cytoplasm of mast cells is filled with many coarse water-soluble, basophilic and heterochromatic granules containing active substances such as dopamine, histamine, heparin, and 5-hydroxytryptamine. These substances contain polysulfated lipids and sulfated mucopolysaccharides, which can be combined with heterochromatic dyes. Therefore, treatment of epithelial tissues with dyes such as Neutral Red, American Blue, and Toluidine Blue can reveal the water-soluble, basophilic, and heterochromatic granules in the cytoplasm of mast cells.

Materials and Instruments

Adult mice
A-F fixative Toluidine blue staining solution Glacial acetic acid color separation solution
Microscope Dissecting scissors Dissecting forceps Used needles Dissecting trays Carriers Covers

Move

1. Preparation of subcutaneous tissue sheets in mice

(1) Neck-breaking method to execute mice

(2) Dissection of mice to remove connective tissue

(3) Spread the tissue on a clean slide and peel the tissue with a used needle, the thinner the better, the size of 0.5cm×0.5cm is sufficient.

(4) Dry the tissue naturally.

2. Toluidine blue staining shows mast cells

(1) Fixation: take the dried sheet and fix it in A-F fixative for 5min.

(2) Rinse: take the fixed slice and rinse it in fine water for 1min.

(3) Staining: put the rinsed slides into 0.5% toluidine blue staining solution for 30 min.

(4) Rinsing: Take the stained slide and rinse it in fine water for 2min.

(5) Color separation: put into 1% glacial acetic acid for a few seconds.

(6) Rinse: take the color-separated film and rinse it in fine water for 2min.

(7) Microscopic examination: microscopic observation (dehydrated, transparent, neutral adhesive sealing film after the better effect)

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