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Method for performing a lymphocyte transformation test by flow cytometry

机译:通过流式细胞术进行淋巴细胞转化试验的方法

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.
机译:进行刺激性淋巴细胞转化试验的方法,包括获取无菌的肝素化样本,将全血或分离的淋巴细胞植入培养基中,在无菌培养箱中孵育,建立两种培养物,其中一种培养物必须包含如下浓度的刺激物:产生细胞增殖,其特征在于在25至45°C的温度下孵育24至192小时,并且包括以下操作的组合: 在培养结束时进行离心; 重新悬浮在红细胞裂解液中; DNA染色; 用等渗水溶液洗涤白细胞悬液; 离心得到的白细胞悬液; 将细胞重新悬浮在裂解白细胞的溶液中在存在DNA特异性荧光染料的情况下,裂解液还可以选择包含用于优化裂解的试剂; 使用流式细胞仪分析样品。

著录项

  • 公开/公告号EP0596840A1

    专利类型

  • 公开/公告日1994-05-11

    原文格式PDF

  • 申请/专利权人 BUSSA STEFANO;RUMI CARLO;

    申请/专利号EP19930830426

  • 发明设计人 BUSSA STEFANO;RUMI CARLO;

    申请日1993-10-26

  • 分类号G01N33/50;G01N33/569;

  • 国家 EP

  • 入库时间 2022-08-22 04:38:59

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