Wang X, Ghosh S, Guo S W
Max McGee National Research Center for Juvenile Diabetes, Medical College and Children's Hospital of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA.
Nucleic Acids Res. 2001 Aug 1;29(15):E75-5. doi: 10.1093/nar/29.15.e75.
A new integrated image analysis package with quantitative quality control schemes is described for cDNA microarray technology. The package employs an iterative algorithm that utilizes both intensity characteristics and spatial information of the spots on a microarray image for signal-background segmentation and defines five quality scores for each spot to record irregularities in spot intensity, size and background noise levels. A composite score q(com) is defined based on these individual scores to give an overall assessment of spot quality. Using q(com) we demonstrate that the inherent variability in intensity ratio measurements is closely correlated with spot quality, namely spots with higher quality give less variable measurements and vice versa. In addition, gauging data by q(com) can improve data reliability dramatically and efficiently. We further show that the variability in ratio measurements drops exponentially with increasing q(com) and, for the majority of spots at the high quality end, this improvement is mainly due to an improvement in correlation between the two dyes. Based on these studies, we discuss the potential of quantitative quality control for microarray data and the possibility of filtering and normalizing microarray data using a quality metrics-dependent scheme.
本文介绍了一种用于cDNA微阵列技术的、带有定量质量控制方案的新型集成图像分析软件包。该软件包采用了一种迭代算法,利用微阵列图像上斑点的强度特征和空间信息进行信号-背景分割,并为每个斑点定义五个质量分数,以记录斑点强度、大小和背景噪声水平的不规则性。基于这些个体分数定义了一个综合分数q(com),以给出斑点质量的总体评估。使用q(com),我们证明了强度比测量中的固有变异性与斑点质量密切相关,即质量较高的斑点测量变异性较小,反之亦然。此外,通过q(com)评估数据可以显著且有效地提高数据可靠性。我们进一步表明,随着q(com)的增加,比率测量的变异性呈指数下降,并且对于高质量端的大多数斑点,这种改进主要是由于两种染料之间相关性的提高。基于这些研究,我们讨论了微阵列数据定量质量控制的潜力,以及使用依赖质量指标的方案对微阵列数据进行过滤和归一化的可能性。