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Measuring de-granulation of basophiles by flow cytometry - after staining leucocytes with Alcian blue, useful in diagnosing allergies
Measuring de-granulation of basophiles by flow cytometry - after staining leucocytes with Alcian blue, useful in diagnosing allergies
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机译：通过流式细胞仪测量嗜碱性粒细胞的脱颗粒-用阿尔辛蓝对白细胞染色后，对诊断过敏有用
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摘要
Method for differential counting of blood basophiles in a control sample and in a sample treated with a degranulating agent, uses flow cytometry after staining with Alcian Blue (I), the degranulation and staining being carried out on the leucocytes after sepn. of cells from plasma and haemolysis of erythrocytes. Pref. a blood sample is treated with heparin, sedimented using a dextrin soln. and the cells centrifuged off. These are suspended in a buffer contg. NH4Cl (to effect haemolysis), then washed with a buffer contg. albumin as stabiliser, and resuspended in the same buffer also contg. Ca and Mg salts to facilitate degranulation. This suspension is treated with the degranulating agent under test (or with an equal vol. of buffer), incubated at 37 deg.C for at least 15 mins. to complete degranulation, then the cells washed with buffer contg. a chelating agent. They are then resuspended in buffer at a concn. of 3-4 million cells per ml, stained with (I) and after 10 mins. counted to determine the no. of basophiles per 100000 leucocytes. The method is used for diagnosis of allergies; allergens will induce a greater degree of degranulation in a sensitive person than in one who is not allergic.
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