磨砂和光滑载玻片对彗星实验检测的影响

摘要

In this paper,the effects of different glass slides（frost and smooth） on the sensitivity and accuracy of the comet assay had been evaluated.The ultraviolet（UV） radiation was used to induce the DNA damage in K562 cell for 3 min and 6min,which was been detected by the comet assay.When carrying out the comet assay,the frosted and conventional glass slides were used separately.The results showed that when using the conventional smooth slide,the background was very dark and clear;but when using the frosted slide the background was brighter and had many bright spots.The tail length was 54.83±16.22 pix and 60.05±17.25 pix when the DNA damage induced by UV for 3 min was detected separately with frosted and conventional smooth slide used in comet assay,and the difference is significant（P0.05）.And the tail length was 62.73±15.29 pix and 74.96±15.67 pix when the DNA damage was induced by UV for 6 min,and the difference is more significant（P0.01）.These results indicated that the frosted glass slide may decrease the sensitivity of comet assay,and the conventional smooth slide should be chosen in the comet assay.%比较磨砂载玻片和常规光滑载玻片对彗星实验结果的影响,以筛选最佳彗星实验用玻片。分别采用磨砂载玻片和常规载玻片进行彗星实验,检测紫外线处理K562细胞3 min和6 min诱导的DNA损伤,采用CASP分析软件比较两种不同载玻片检测的灵敏度。结果表明,常规载玻片的背景荧光较弱,而磨砂载玻片背景荧光较强,且有斑点状杂质荧光;软件分析结果显示照射3 min时,分别采用磨砂和常规载玻片检测到的尾长为54.83±16.22像素和60.05±17.25像素;照射6 min时的尾长分别为62.73±15.29像素和74.96±15.67像素。结果显示磨砂载玻片影响图像的采集和分析,降低了彗星实验检测的灵敏度,提示在彗星实验中应选择常规光滑载玻片进行实验。