Gardell D, Rock G
Canadian Red Cross Society, Blood Transfusion Service Ottawa Center, Ontario.
Lab Invest. 1988 May;58(5):599-606.
Automated contrast fluorometry has been carried out in lymphocytotoxicity testing using a computer-controlled microscope and direct comparison of the data with controls. The method uses heparinized lymphocytes and permits simultaneous evaluation of both fluorescein and ethidium bromide fluorescence by calculating the quotient between the green/red measurements. After comparison of the quotient with that of control samples, the raw data are automatically transformed into cytotoxicity scores. Fast and reliable measurements with minimum background fluorescence can be obtained for up to 24 hours after performance of the assay by controlling complement reactivity and the exposure time of the cells to the fluorescein. The evaluation of 15,204 cytotoxic reactions showed excellent correlation between the automated and visual results of 125 tissue typings using 120 different antibodies and of 17 HLA antibody titers using case-by-case analysis of variance. The values for the automated cytotoxicity readings were slightly higher and showed more intermediate scores than those of the visual readings suggesting a high sensitivity and objectivity of the automated system. In conclusion, this is a methods paper comparing parallel lymphocytotoxicity testing using automated and manual methods. The automated method was found to be more sensitive and faster than the manual method.
在淋巴细胞毒性检测中，使用计算机控制的显微镜进行自动对比荧光测定，并将数据与对照进行直接比较。该方法使用肝素化淋巴细胞，通过计算绿色/红色测量值之间的商，同时评估荧光素和溴化乙锭的荧光。将该商数与对照样品的商数进行比较后，原始数据会自动转换为细胞毒性评分。通过控制补体反应性和细胞与荧光素的接触时间，在检测完成后的长达24小时内，可以获得背景荧光最小的快速可靠测量结果。对15204次细胞毒性反应的评估表明，在使用120种不同抗体的125次组织分型以及使用逐例方差分析的17种HLA抗体滴度的自动和视觉结果之间，具有极好的相关性。自动细胞毒性读数的值略高，并且与视觉读数相比显示出更多的中间分数，这表明自动系统具有高灵敏度和客观性。总之，这是一篇比较使用自动和手动方法进行平行淋巴细胞毒性检测的方法论文。发现自动方法比手动方法更灵敏、更快。
