MOTIVATION: A serious limitation in microarray analysis is the unreliability of the data generated from low signal intensities. Such data may produce erroneous gene expression ratios and cause unnecessary validation or post-analysis follow-up tasks. Therefore, the elimination of unreliable signal intensities will enhance reproducibility and reliability of gene expression ratios produced from microarray data. In this study, we applied fuzzy c-means (FCM) and normal mixture modeling (NMM) based classification methods to separate microarray data into reliable and unreliable signal intensity populations. RESULTS: We compared the results of FCM classification with those of classification based on NMM. Both approaches were validated against reference sets of biological data consisting of only true positives and true negatives. We observed that both methods performed equally well in terms of sensitivity and specificity. Although a comparison of the computation times indicated that the fuzzy approach is computationally more efficient, other considerations support the use of NMM for the reliability analysis of microarray data. AVAILABILITY: The classification approaches described in this paper and sample microarray data are available as Matlab( TM ) (The MathWorks Inc., Natick, MA) programs (mfiles) and text files, respectively, at http://rc.kfshrc.edu.sa/bssc/staff/MusaAsyali/Downloads.asp. The programs can be run/tested on many different computer platforms where Matlab is available. CONTACT: [email protected].

MOTIVATION: Spot segmentation is a critical step in microarray gene expression data analysis. Therefore, the performance of segmentation may substantially affect the results of subsequent stages of the analysis, such as the detection of differentially expressed genes. Several methods have been developed to segment microarray spots from the surrounding background. In this study, we have proposed a new approach based on Markov random field (MRF) modeling and tested its performance on simulated and real microarray images against a widely used segmentation method based on Mann-Whitney test adopted by QuantArray software (Boston, MA). Spot addressing was performed using QuantArray. We have also devised a simulation method to generate microarray images with realistic features. Such images can be used as gold standards for the purposes of testing and comparing different segmentation methods, and optimizing segmentation parameters. RESULTS: Experiments on simulated and 14 actual microarray image sets show that the proposed MRF-based segmentation method can detect spot areas and estimate spot intensities with higher accuracy.