An Automated cDNA Microarray Image Analysis for the Determination of Gene Expression Ratios.

Abstract

This paper proposes a fully automated technique for cDNA microarray image analysis. Initially, an effective preprocessing stage combined with gridding is built to get the individual spot regions of images. Current work begins with the proposal of a new rule to get the foreground (spot) and background regions in the spot blocks, which uses TV-L1 image denoising, spot block binarization, and finds the most accurate spot label by measuring the centroid differences of labelled regions in the block with that of the spot block centroid. The credibility of the segmentation rule on real images is evaluated by metrics: mean absolute error (MAE) and coefficient of variation (CV) and on synthetic images by metrics: probability of error (PE) and discrepancy distance (DD). The performance values on real and synthetic datasets reveal better results than the competitive methods. After the segmentation, prior to the spot intensity extraction, background intensity correction and flagging of noisy spots are executed. Using the lowess method, intensities are normalized, and gene expression ratios are determined. To comprehend the linearities of red and green intensities and to discern up and down-regulated genes (abnormal), fold-change factor, scatter and box plots are also used to represent the gene expression levels.