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Method for performing a lymphocyte transformation test by flow cytometry
Method for performing a lymphocyte transformation test by flow cytometry
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机译:通过流式细胞术进行淋巴细胞转化试验的方法
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摘要
Method for performing a stimulated lymphocyte transformation test, comprising taking a sterile heparinated sample, implanting the whole blood or separated lymphocytes in culture medium, incubation in a sterile incubator, setting up two cultures, one of which must contain the stimulant at concentrations such as to generate cell proliferation, characterized by the fact that incubation takes place at a temperature of between 25 and 45°C for a time of between 24 and 192 hours, and by the fact that it comprises the combination of the following operations:centrifugation at the end of the incubation;re-suspension in a lysing solution for erythrocytes;DNA staining;washing the leucocyte suspension with isotonic aqueous solution;centrifugation of the resulting leucocyte suspension;re-suspension of the cells in a solution lysing for leucocytes, in the presence of a DNA-specific fluorochrome, the lysing solution also optionally comprising reagents for optimization of lysis;analysis of the samples, using flow cytometer.展开▼