Automated cDNA Microarray Image Segmentation

Using fuzzy logic and particle swarm optimization to design a decision-based filter for cDNA microarray image restoration

Gridding and spot segmentation are two critical steps in microarray gene expression data analysis. However, the problems of noise contamination and donut-shaped spots often make signal extraction process a laborintensive task. In this paper, we propose a three-step method for automatic gridding and spot segmentation. The method starts with a background removal and noise eliminating step, and then proceeds in two steps. The first step applies a fully unsupervised method to extract blocks and grids from the cleaned data. The second step applies a simple, progressive spot segmentation method to deal with inner holes and noise in spots. We tested its performance on real microarray images against a widely used software GenePix. Our results show that the proposed method deals effectively with poor-conditioned microarray images in both gridding and spot segmentation.

Background/Objectives: This study presents an external and internal prior guided patch based filter for minimizing Gaussian noise in complementary deoxyribonucleic acid (cDNA) microarray images. Methods/Statistical analysis: The proposed denoising filter is capable of taking into consideration both external and internal prior. It employs combined prior guided patch based denoising considering various distance based patch-matching methods. Findings: Experimental results demonstrate that the combined prior guided patch based filter outperforms the existing well-known filters in minimizing noise of cDNA microarray images. The outcome of the proposed scheme found to offer better peak signalto-noise ratio and structural similarity index in contrast to existing filtering techniques. Effectiveness of the proposed denoising method is also assessed by estimating the spot intensities of cDNA microarray image that reflects the effect of noise reduction in the image. Application/Improvements: Minimization of noise is a crucial step of cDNA microarray image processing and it aids in microarray analysis by extracting valid and good quality gene expression measurements. Keywords: Denoising, External Prior, Internal Prior, K- nearest Neighbor, Microarray Image, Singular Value Decomposition

DNA microarray technology has permitted the analysis of global gene expression profiles in clinical diagnosis, treatment, and environmental health research for several diseases including cancer. Microarray images analysis, processing and segmentation are decisive steps in gene expression analysis, since any error leads to improper diagnosis. These techniques increased and enhanced over the past few years. However, the segmentation of the unshaped spots within the microarray image is still a problem due to the variations of spots in shapes and sizes. Hence, we introduce a new unshaped microarray spot segmentation model, which focuses on spots detection and segmentation regardless of their size and shape using fractal dimensions transform. Real microarray images from the Stanford Microarray Images Database (SMD) and Universal Microarray Images Database (UNC) are used to evaluate the efficiency of the proposed model. The numerical and visual results show that the proposed model improves the accuracy and the efficiency of the spots segmentation process. Biological researchers can use this model in their labs and clinics, as a free priceless software instead of using expensive ones like ScanAlyze, GenePix or spot software.

The accuracy of gene expression measurements generated using cDNA microarrays is dependent on the quality of the image generated following hybridization of fluorescently labelled cDNA. It is not known how this image is influenced by sample preparation factors which such as RNA quality, cDNA synthesis and labelling efficiency. In this study we used a simple metric based on the ratio of the total feature (F) and background (B) fluorescence, which correlates with the visual assessment of 60 microarray images, to determine the influence of sample preparation on image quality. Results indicate that RNA purity (A260/A280) and integrity (18S:28S ratio) do not strongly influence microarray image quality. cDNA having an nucleotide to dye ratio greater than 100 produced poor microarray images, however, cDNA labelled more efficiently was not a guarantee of a better image. The data also indicate that the array image quality is not improved by loading more cDNA into the hybridization mixture however poor image quality did result from a disproportionate amounts of Cy5 and Cy3 labelled cDNA. This study provides insight into the source of variation in microarray image analysis introduced during sample preparation and will assist in the standardisation of cDNA glass slide microarray protocols.

Complementary DNA microarray technology is a powerful tool in many areas. Usually a two channel microarray red-green (RG) image is obtained. Due to the nature of cDNA microarray technology, a number of impairments affect the cDNA microarray image before the analysis such as identification of differentially expressed genes. Microarray image processing plays a crucial role in the extraction and quantitative analysis of the relative abundance of the DNA product. In this paper, a method combined K-means clustering algorithm and mathematical morphology is presented. Mathematical morphology is a useful tool for extracting image components. K-means clustering algorithm has a good performance in the segmentation of microarray image processing. The result of the experiment shows that the method presented in this paper is accurate, automatic and robust.

A wavelet-based Markov random field segmentation model in segmenting microarray experiments

Microarray technology has become a power tool in the field of bioinformatics. It is used to measure gene expression levels and similar to any other image capturing processes is prone to noise. There are different kinds of noise, during preparation, hybridization and scanning in microarray images which usually are modeled by Gaussian noise. Since introduction of wavelets in 1970s, many more forms and extensions of this transform have been developed and used, such as stationary wavelet transform (SWT), complex wavelet transform (CWT), curvelet transform (CURV) and contourlet transform (CNT). By developing of more sparse transforms, it is important to have a perspective of how efficient the transforms are in different applications, such as microarray image analysis. In this paper, we compare the efficiency of common sparse transforms including ordinary discrete wavelet transform (DWT), SWT, CWT, CURV, CNT, Contourlet-SD decomposition, steerable pyramid (STP) and shearlet transform (SHR) for microarray image denoising. Therefore after converting microarray image into x-let transform, BayesShrink method, soft and hard thresholding are used to perform denoising of these images. Both local and general thresholds are calculated for each subband in order to evaluate the effect of incorporating intrascale dependency on top of sparsity property in statistical modeling of x-let's coefficients. Our simulation results show that CWT and SHR outperforms the others when using global thresholding and SWT is the preferred transform when using local thresholding. Although STP and SHR have better performance for some criteria like structural similarity (SSIM) index, but CWT is faster.

Image processing is an important stage of every microarray experiment. Reliability of this stage strongly influences the results of data analysis performed on extracted gene expressions. Multiple methods related to array recognition, spot segmentation and measurement extraction have emerged in this area over past several years. Currently there are various commercial and freeware packages available, which perform microarray image analysis. This paper attempts to review microarray image analysis as a whole and to make some experimental comparison of several computational schemes for signal segmentation and measurement extraction. Also we provide a detailed discussion of automated image quality control for use with microarray images.

Microarray images, which allow the analysis of hybridization expressions of genes, have been one of the most widely used high-throughput technologies with many different applications. Accurate and automatic microarray image analysis is very important since researchers trust the information provided in these experiments and construct conclusions based on the results produced by the software responsible in analyzing the hybridized arrays. Every microarray image contains thousands of spots, so how to do the spots finding in microarray images accurately and automatically is very meaningful. There are always some problems, such as rotation and distortion, in a microarray image caused by mechanical errors and/or optical errors in the system. All these problems will hinder "doi"ng analysis automatically. Early research has worked out several algorithms to deal with the rotation problem, but those algorithms can not handle microarray images with distortions. In this paper, we propose a robust spots finding method to deal with both rotation and/or distortion in microarray images. The proposed method provides automatic gridding and can handle a microarray image with different type of rotation (global or sub-array rotation) and optical distortions.

An automated method for gridding and clustering-based segmentation of cDNA microarray images

Genetic microarray image processing has become an important field of research to produce tools aiming to reliable gene expression estimation. Current instrumentation to analyze microarray images lacks flexibility and accuracy. In the present paper an FPGA-based hand-held device for the analysis of microarrays is described, which may implement basic scanning and reticle algorithms to estimate spots and expression levels in cDNA microarrays. The algorithms are based on global and local spot correlation, background removal and rectangular mask fitting. These algorithms can be recursively applied and are intrinsically parallelizable for practical implementation purposes. Results using specific microarray images are shown.

BackgroundThe quality of cDNA microarray data is crucial for expanding its application to other research areas, such as the study of gene regulatory networks. Despite the fact that a number of algorithms have been suggested to increase the accuracy of microarray gene expression data, it is necessary to obtain reliable microarray images by improving wet-lab experiments. As the first step of a cDNA microarray experiment, spotting cDNA probes is critical to determining the quality of spot images.ResultsWe developed a governing equation of cDNA deposition during evaporation of a drop in the microarray spotting process. The governing equation included four parameters: the surface site density on the support, the extrapolated equilibrium constant for the binding of cDNA molecules with surface sites on glass slides, the macromolecular interaction factor, and the volume constant of a drop of cDNA solution. We simulated cDNA deposition from the single model equation by varying the value of the parameters. The morphology of the resulting cDNA deposit can be classified into three types: a doughnut shape, a peak shape, and a volcano shape. The spot morphology can be changed into a flat shape by varying the experimental conditions while considering the parameters of the governing equation of cDNA deposition. The four parameters were estimated by fitting the governing equation to the real microarray images. With the results of the simulation and the parameter estimation, the phenomenon of the formation of cDNA deposits in each type was investigated.ConclusionThis study explains how various spot shapes can exist and suggests which parameters are to be adjusted for obtaining a good spot. This system is able to explore the cDNA microarray spotting process in a predictable, manageable and descriptive manner. We hope it can provide a way to predict the incidents that can occur during a real cDNA microarray experiment, and produce useful data for several research applications involving cDNA microarrays.

Microarray image compression: SLOCO and the effect of information loss

Current gene-expression microarrays carry enormous amounts of information. Compression is necessary for efficient distribution and storage. This paper examines JPEG2000 compression of cDNA microarray images and addresses the accuracy of classification and feature selection based on decompressed images. Among other options, we choose JPEG2000 because it is the latest international standard for image compression and offers lossy-to-lossless compression while achieving high lossless compression ratios on microarray images. The performance of JPEG2000 has been tested on three real data sets at different compression ratios, ranging from lossless to 45:1. The effects of JPEG2000 compression/decompression on differential expression detection and phenotype classification have been examined. There is less than a 4% change in differential detection at compression rates as high as 20:1, with detection accuracy suffering less than 2% for moderate to high intensity genes, and there is no significant effect on classification at rates as high as 35:1. The supplementary material is available at http://gsp.tamu.edu/web2/Compression.