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首页> 外文期刊>Mutation Research. Reviews in Mutation Research >Some causes of inter-laboratory variation in the results of comet assay
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Some causes of inter-laboratory variation in the results of comet assay

机译：彗星试验结果实验室间变异的一些原因

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We performed an inter-laboratory study to determine the variation of comet assay results and to identify its possible reasons. An exchange of slides between Labs in different stages of the comet assay protocol was performed. Because identical slides, durations of alkali treatment and electrophoresis, and similar electric field strengths (2.0V/cm and 2.14V/cm) were used, we concluded that the observed inter-laboratory difference in the results is directly associated with the electrophoresis step. In Lab 1, mouse bone marrow cells were exposed to methyl methanesulfonate at concentrations of 10,25 and 50 fxM for 3 h at 37 °C. In Lab 2, cells the same as in Lab 1 were immobilized in LMA on slides and exposed to X-rays at doses of 3-8 Gy. We found that the transportation of slides after lysis or electrophoresis step, as well as different dyes used for scoring did not produce any significant effect on the results. No substantial difference in the data was also revealed when various software packages were used for image analysis. The temperature of the alkaline solution was shown to increase during electrophoresis and, besides, the temperature heterogeneity of the solution took place in the area of the platform, with a maximum in the middle of the chamber. The temperature heterogeneity could affect the rate of conversion of alkali labile sites into single stranded breaks. Thus, it was clearly indicated that real temperature variations during the alkali treatment and electrophoresis were an essential factor in the variability of the results between our Labs.
机译：我们进行了一项实验室间研究，以确定彗星分析结果的变化并确定其可能的原因。在彗星试验方案不同阶段的实验室之间进行了载玻片交换。因为使用了相同的载玻片，碱处理和电泳的时间以及相似的电场强度（2.0V / cm和2.14V / cm），所以我们得出的结论是，观察到的实验室间差异与电泳步骤直接相关。在实验室1中，小鼠骨髓细胞在37°C下暴露于浓度为10,25和50 fxM的甲磺酸甲酯中3小时。在实验室2中，将与实验室1中相同的细胞固定在载玻片上的LMA中，并以3-8 Gy的剂量暴露于X射线。我们发现，裂解或电泳步骤后载玻片的运输以及用于评分的不同染料对结果均未产生任何重大影响。当使用各种软件包进行图像分析时，数据也没有显示出实质性差异。碱性溶液的温度在电泳过程中显示出升高，此外，溶液的温度异质性在平台区域发生，最高出现在反应室中间。温度异质性可能影响碱不稳定位点转化为单链断裂的速率。因此，可以清楚地表明，碱处理和电泳过程中的实际温度变化是影响我们实验室之间结果差异的重要因素。

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