A novel neural network approach to cDNA microarray image segmentation

This is the post-print version of the Article. The official published version can be accessed from the link below. Copyright @ 2013 Elsevier.Microarray technology has become a great source of information for biologists to understand the workings of DNA which is one of the most complex codes in nature. Microarray images typically contain several thousands of small spots, each of which represents a different gene in the experiment. One of the key steps in extracting information from a microarray image is the segmentation whose aim is to identify which pixels within an image represent which gene. This task is greatly complicated by noise within the image and a wide degree of variation in the values of the pixels belonging to a typical spot. In the past there have been many methods proposed for the segmentation of microarray image. In this paper, a new method utilizing a series of artificial neural networks, which are based on multi-layer perceptron (MLP) and Kohonen networks, is proposed. The proposed method is applied to a set of real-world cDNA images. Quantitative comparisons between the proposed method and commercial software GenePix(®) are carried out in terms of the peak signal-to-noise ratio (PSNR). This method is shown to not only deliver results comparable and even superior to existing techniques but also have a faster run time.This work was funded in part by the National Natural Science Foundation of China under Grants 61174136 and 61104041, the Natural Science Foundation of Jiangsu Province of China under Grant BK2011598, the International Science and Technology Cooperation Project of China under Grant No. 2011DFA12910, the Engineering and Physical Sciences Research Council (EPSRC) of the U.K. under Grant GR/S27658/01, the Royal Society of the U.K., and the Alexander von Humboldt Foundation of Germany

A fully automatic gridding method for cDNA microarray images

<p>Abstract</p> <p>Background</p> <p>Processing cDNA microarray images is a crucial step in gene expression analysis, since any errors in early stages affect subsequent steps, leading to possibly erroneous biological conclusions. When processing the underlying images, accurately separating the sub-grids and spots is extremely important for subsequent steps that include segmentation, quantification, normalization and clustering.</p> <p>Results</p> <p>We propose a parameterless and fully automatic approach that first detects the sub-grids given the entire microarray image, and then detects the locations of the spots in each sub-grid. The approach, first, detects and corrects rotations in the images by applying an affine transformation, followed by a polynomial-time optimal multi-level thresholding algorithm used to find the positions of the sub-grids in the image and the positions of the spots in each sub-grid. Additionally, a new validity index is proposed in order to find the correct number of sub-grids in the image, and the correct number of spots in each sub-grid. Moreover, a refinement procedure is used to correct possible misalignments and increase the accuracy of the method.</p> <p>Conclusions</p> <p>Extensive experiments on real-life microarray images and a comparison to other methods show that the proposed method performs these tasks fully automatically and with a very high degree of accuracy. Moreover, unlike previous methods, the proposed approach can be used in various type of microarray images with different resolutions and spot sizes and does not need any parameter to be adjusted.</p

Bioinformatics framework for genotyping microarray data analysis

Functional genomics is a flourishing science enabled by recent technological breakthroughs in high-throughput instrumentation and microarray data analysis. Genotyping microarrays establish the genotypes of DNA sequences containing single nucleotide polymorphisms (SNPs), and can help biologists probe the functions of different genes and/or construct complex gene interaction networks. The enormous amount of data from these experiments makes it infeasible to perform manual processing to obtain accurate and reliable results in daily routines. Advanced algorithms as well as an integrated software toolkit are needed to help perform reliable and fast data analysis. The author developed a MatlabTM based software package, called TIMDA (a Toolkit for Integrated Genotyping Microarray Data Analysis), for fully automatic, accurate and reliable genotyping microarray data analysis. The author also developed new algorithms for image processing and genotype-calling. The modular design of TIMDA allows satisfactory extensibility and maintainability. TIMDA is open source (URL: http://timda.SF.net and can be easily customized by users to meet their particular needs. The quality and reproducibility of results in image processing and genotype-calling and the ease of customization indicate that TIMDA is a useful package for genomics research

Fully automatic classification of breast cancer microarray images

AbstractA microarray image is used as an accurate method for diagnosis of cancerous diseases. The aim of this research is to provide an approach for detection of breast cancer type. First, raw data is extracted from microarray images. Determining the exact location of each gene is carried out using image processing techniques. Then, by the sum of the pixels associated with each gene, the amount of “genes expression” is extracted as raw data. To identify more effective genes, information gain method on the set of raw data is used. Finally, the type of cancer can be recognized via analyzing the obtained data using a decision tree. The proposed approach has an accuracy of 95.23% in diagnosing the breast cancer types

M3G: Maximum Margin Microarray Gridding

<p>Abstract</p> <p>Background</p> <p>Complementary DNA (cDNA) microarrays are a well established technology for studying gene expression. A microarray image is obtained by laser scanning a hybridized cDNA microarray, which consists of thousands of spots representing chains of cDNA sequences, arranged in a two-dimensional array. The separation of the spots into distinct cells is widely known as microarray image gridding.</p> <p>Methods</p> <p>In this paper we propose M³G, a novel method for automatic gridding of cDNA microarray images based on the maximization of the margin between the rows and the columns of the spots. Initially the microarray image rotation is estimated and then a pre-processing algorithm is applied for a rough spot detection. In order to diminish the effect of artefacts, only a subset of the detected spots is selected by matching the distribution of the spot sizes to the normal distribution. Then, a set of grid lines is placed on the image in order to separate each pair of consecutive rows and columns of the selected spots. The optimal positioning of the lines is determined by maximizing the margin between these rows and columns by using a maximum margin linear classifier, effectively facilitating the localization of the spots.</p> <p>Results</p> <p>The experimental evaluation was based on a reference set of microarray images containing more than two million spots in total. The results show that M³G outperforms state of the art methods, demonstrating robustness in the presence of noise and artefacts. More than 98% of the spots reside completely inside their respective grid cells, whereas the mean distance between the spot center and the grid cell center is 1.2 pixels.</p> <p>Conclusions</p> <p>The proposed method performs highly accurate gridding in the presence of noise and artefacts, while taking into account the input image rotation. Thus, it provides the potential of achieving perfect gridding for the vast majority of the spots.</p

Estimating Gene Signals From Noisy Microarray Images

In oligonucleotide microarray experiments, noise is a challenging problem, as biologists now are studying their organisms not in isolation but in the context of a natural environment. In low photomultiplier tube (PMT) voltage images, weak gene signals and their interactions with the background fluorescence noise are most problematic. In addition, nonspecific sequences bind to array spots intermittently causing inaccurate measurements. Conventional techniques cannot precisely separate the foreground and the background signals. In this paper, we propose analytically based estimation technique. We assume a priori spot-shape information using a circular outer periphery with an elliptical center hole. We assume Gaussian statistics for modeling both the foreground and background signals. The mean of the foreground signal quantifies the weak gene signal corresponding to the spot, and the variance gives the measure of the undesired binding that causes fluctuation in the measurement. We propose a foreground-signal and shapeestimation algorithm using the Gibbs sampling method. We compare our developed algorithm with the existing Mann–Whitney (MW)- and expectation maximization (EM)/iterated conditional modes (ICM)-based methods. Our method outperforms the existing methods with considerably smaller mean-square error (MSE) for all signal-to-noise ratios (SNRs) in computer-generated images and gives better qualitative results in low-SNR real-data images. Our method is computationally relatively slow because of its inherent sampling operation and hence only applicable to very noisy-spot images. In a realistic example using our method, we show that the gene-signal fluctuations on the estimated foreground are better observed for the input noisy images with relatively higher undesired bindings

Automatic gridding of DNA microarray images.

Microarray (DNA chip) technology is having a significant impact on genomic studies. Many fields, including drug discovery and toxicological research, will certainly benefit from the use of DNA microarray technology. Microarray analysis is replacing traditional biological assays based on gels, filters and purification columns with small glass chips containing tens of thousands of DNA and protein sequences in agricultural and medical sciences. Microarray functions like biological microprocessors, enabling the rapid and quantitative analysis of gene expression patterns, patient genotypes, drug mechanisms and disease onset and progression on a genomic scale. Image analysis and statistical analysis are two important aspects of microarray technology. Gridding is necessary to accurately identify the location of each of the spots while extracting spot intensities from the microarray images and automating this procedure permits high-throughput analysis. Due to the deficiencies of the equipment that is used to print the arrays, rotations, misalignments, high contaminations with noise and artifacts, solving the grid segmentation problem in an automatic system is not trivial. The existing techniques to solve the automatic grid segmentation problem cover only limited aspect of this challenging problem and requires the user to specify or make assumptions about the spotsize, rows and columns in the grid and boundary conditions. An automatic gridding and spot quantification technique is proposed, which takes a matrix of pixels or a microarray image as input and makes no assumptions about the spotsize, rows and columns in the grid and is found to effective on datasets from GEO, Stanford genomic laboratories and on images obtained from private repositories. Paper copy at Leddy Library: Theses & Major Papers - Basement, West Bldg. / Call Number: Thesis2004 .V53. Source: Masters Abstracts International, Volume: 43-03, page: 0891. Adviser: Luis Rueda. Thesis (M.Sc.)--University of Windsor (Canada), 2004

Automatic gridding of microarray images based on spatial constrained K-means and Voronoi diagrams

Images from complementary DNA (cDNA) microarrays need to be processed automatically due to the huge amount of information that they provide. In addition, automatic processing is also required to implement batch processes able to manage large image databases. Most of existing softwares for microarray image processing are semiautomatic, and they usually need user intervention to select several parameters such as positional marks on the grids, or to correct the results of different stages of the automatic processing. On the other hand, many of the available automatic algorithms fail when dealing with rotated images or misaligned grids. In this work, a novel automatic algorithm for cDNA image gridding based on spatial constrained K-means and Voronoi diagrams is presented. The proposed algorithm consists of several steps, viz., image denoising by means of median filtering, spot segmentation using Canny edge detector and morphological reconstruction, and gridding based on spatial constrained K-means and Voronoi diagrams computation. The performance of the algorithm was evaluated on microarray images from public databases yielding promising results. The algorithm was compared with other existing methods and it shows to be more robust to rotations and misalignments of the grids.Red de Universidades con Carreras en Informática (RedUNCI

Automating the processing of cDNA microarray images

This work is concerned with the development of an automatic image processing tool for DNA microarray images. This paper proposes, implements and tests a new tool for cDNA image analysis. The DNAs are imaged as thousands of circularly shaped objects (spots) on the microarray image and the purpose of this tool is to correctly address their location, segment the pixels belonging to spots and extract the quality features of each spot. Techniques used for addressing, segmentation and feature extraction of spots are described in detail. The results obtained with the proposed tool are systematically compared with conventional cDNA microarray analysis software tools

An Overview of DNA Microarray Grid Alignment and Foreground Separation Approaches

This paper overviews DNA microarray grid alignment and foreground separation approaches. Microarray grid alignment and foreground separation are the basic processing steps of DNA microarray images that affect the quality of gene expression information, and hence impact our confidence in any data-derived biological conclusions. Thus, understanding microarray data processing steps becomes critical for performing optimal microarray data analysis. In the past, the grid alignment and foreground separation steps have not been covered extensively in the survey literature. We present several classifications of existing algorithms, and describe the fundamental principles of these algorithms. Challenges related to automation and reliability of processed image data are outlined at the end of this overview paper.</p