显微图像分析系统测量细胞核DNA含量误差的研究

Measurement errors of nuclear DNA contents with a digital imaging microscopy system

检测显微图像分析系统测量细胞核ＤＮＡ含量的误差对于判断肿瘤细胞ＤＮＡ倍体分析的准确性具有重要意义。本文检测了ＴＩＧＥＲ显微图像分析仪的系统噪声、聚焦变化、光源亮度与光谱差异、背景光强度分布不均匀的测量误差。结果显示，（１）尽管实现了Ｉｏ和Ｉ在同一空间位置的测量，但是，它们仍给测量结果引入了测量误差，以背景光强度分布不均导致ＩＯＤ的测量误差最大；（２）应根据目标细胞的大小和密度，选择适当的物镜倍率，样品制备应使ＡＯＤ达到一定的密度值以上，才能最大限度地降低测量误差；（３）鸡红细胞核是综合检测和评价显微图像分析系统测量细胞光密度性能的良好生物学标准。

Measurement errors of nuclear DNA content influence the accuracy of DNA ploidy analysis. The purpose of the present study was to analyse factors including system noise, focus and inhomogeneity of background light intensity, that produce influences on measurment errors evaluated by TIGER image analysis system. It is concluded that (1) the inhomogeneity of background light intensity results in the greatest IOD measurement errors and I o and I must be measured at the same local position; (2) the objective should be suitably chosen according to cellular size and AOD which has to be reach some density value by proper staining, in order to minimize measurement errors of IOD, AOD and A; (3) CRBC is the good biologic standarization for checking and assessing the quality and perforemance of the digital imaging microscopy system.