微核试验制片方法的改进

The improvement of micronucleus test method

目的简化、优化微核试验方法。方法对微核试验的制片过程进行简化、优化,改进后在细胞培养结束后直接吸弃上清液,然后加入氯化钾溶液进行低渗处理,随后预固定、离心。离心完成后,细胞再固定一次即可滴片。结果改进法玻片背景清晰,细胞染色稍深,但不影响细胞和微核观察。双核细胞数量不少,能够满足计数要求。油镜和高倍镜下,图像更清楚、背景更干净。改进法胞浆完整率、细胞着色率和平均每高倍视野细胞个数与传统方法比较有统计学意义,概率P值分别为0.0051(χ^(2)=7.8375)、0.0140(χ^(2)=6.0437)和0.0025(t=3.0951)。微核细胞率和细胞成团指数与传统方法比较无统计学意义,概率P值分别为0.7749(χ^(2)=0.0817)和0.5152(U=0.0000)。结论改进法制片方法简单易行、结果可靠,试验质量更容易控制,还节省了时间,节省了人力、物力。

Objective To simplify and optimize the micronucleus test method.Methods The preparation process of micronucleus test was simplified and optimized.In the improved method,the superfine solution was directly absorbed and discarded after cell culture,and then potassium chloride solution was added for hypotonic treatment.Then pre-fixation,centrifugation.Once the centrifugation was completed,the cells which fixed only once were directly dropped to the slide.Results The background of the slides was clear and the cells were slightly darker,but the observation of cells and micronucleus was not affected.There were a lot of binuclear cells,which can meet the counting requirements.With oil and high magnification,the image was clearer and the background was cleaner.The cytoplasmic integrity rate,cell stain rate and the average number of cells per high magnification field of cells by the improvement method were significantly increased compared with that by the traditional methods,the probability P values were 0.0051(χ^(2)=7.8375),0.0140(χ^(2)=6.0437)and 0.0025(t=3.0951),respectively.The rate of micronucleus cells and cells group index had no statistical significance compared with the traditional method,the probability P values were 0.7749(χ^(2)=0.0817)and 0.5152(U=0.0000),respectively.Conclusion The new method is more simple,easier to control the test quality,more reliable test results,and save time,manpower and material resources.