A New Gridding Technique for High Density Microarray Images Using Intensity Projection Profile of Best Sub Image

As the technologies for the fabrication of high quality microarray advances rapidly, quantification of microarray data becomes a major task. Gridding is the first step in the analysis of microarray images for locating the subarrays and individual spots within each subarray. For accurate gridding of high-density microarray images, in the presence of contamination and background noise, precise calculation of parameters is essential. This paper presents an accurate fully automatic gridding method for locating suarrays and individual spots using the intensity projection profile of the most suitable subimage. The method is capable of processing the image without any user intervention and does not demand any input parameters as many other commercial and academic packages. According to results obtained, the accuracy of our algorithm is between 95-100% for microarray images with coefficient of variation less than two.  Experimental results show that the method is capable of gridding microarray images with irregular spots, varying surface intensity distribution and with more than 50% contamination. Keywords: microarray, gridding, image processing, gridding accurac

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

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

Medical Image Segmentation Using Multifractal Analysis

Image segmentation plays a key role in image analysis processes. The operations performed on a segmented image tend to affect it differently than if they were performed on the original image; therefore, segmenting an image can show radically different results from the original image and successfully doing so can yield features and other important information about the image. Proper image analysis is of high importance to the medical community as accurately classifying different conditions and diseases can be facilitated with excellent patient imaging. Multifractal analysis can be leveraged for performing texture classification and image segmentation. In this paper, we propose fusion-based algorithms utilizing multifractal analysis for medical image segmentation. We use two specific multifractal masks: square and quincunx. Our techniques show new insights by using methods such as histogram decomposition in conjunction with new techniques, such as fusion. By fusing different slope images, we can extract more features thus making our proposed algorithms more robust and accurate than traditional multifractal analysis techniques. These methods are further capable of reliably segmenting medical images by implementing multifractal analysis techniques in coordination with methods such as gaussian blurring and morphological operations. The resulting image can then be easily analyzed by medical professionals for diagnosing medical conditions. The outcomes show that the proposed algorithms extract dominant features that are more encompassing and powerful than classical techniques

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

Crossword: A Fully Automated Algorithm for the Segmentation and Quality Control of Protein Microarray Images

Biological assays formatted as microarrays have become a critical tool for the generation of the comprehensive data sets required for systems-level understanding of biological processes. Manual annotation of data extracted from images of microarrays, however, remains a significant bottleneck, particularly for protein microarrays due to the sensitivity of this technology to weak artifact signal. In order to automate the extraction and curation of data from protein microarrays, we describe an algorithm called Crossword that logically combines information from multiple approaches to fully automate microarray segmentation. Automated artifact removal is also accomplished by segregating structured pixels from the background noise using iterative clustering and pixel connectivity. Correlation of the location of structured pixels across image channels is used to identify and remove artifact pixels from the image prior to data extraction. This component improves the accuracy of data sets while reducing the requirement for time-consuming visual inspection of the data. Crossword enables a fully automated protocol that is robust to significant spatial and intensity aberrations. Overall, the average amount of user intervention is reduced by an order of magnitude and the data quality is increased through artifact removal and reduced user variability. The increase in throughput should aid the further implementation of microarray technologies in clinical studies.Camille and Henry Dreyfus Foundation (Camille Dreyfus Teacher-Scholar Award

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

Microarray image analysis: background estimation using quantile and morphological filters

BACKGROUND: In a microarray experiment the difference in expression between genes on the same slide is up to 10(3 )fold or more. At low expression, even a small error in the estimate will have great influence on the final test and reference ratios. In addition to the true spot intensity the scanned signal consists of different kinds of noise referred to as background. In order to assess the true spot intensity background must be subtracted. The standard approach to estimate background intensities is to assume they are equal to the intensity levels between spots. In the literature, morphological opening is suggested to be one of the best methods for estimating background this way. RESULTS: This paper examines fundamental properties of rank and quantile filters, which include morphological filters at the extremes, with focus on their ability to estimate between-spot intensity levels. The bias and variance of these filter estimates are driven by the number of background pixels used and their distributions. A new rank-filter algorithm is implemented and compared to methods available in Spot by CSIRO and GenePix Pro by Axon Instruments. Spot's morphological opening has a mean bias between -47 and -248 compared to a bias between 2 and -2 for the rank filter and the variability of the morphological opening estimate is 3 times higher than for the rank filter. The mean bias of Spot's second method, morph.close.open, is between -5 and -16 and the variability is approximately the same as for morphological opening. The variability of GenePix Pro's region-based estimate is more than ten times higher than the variability of the rank-filter estimate and with slightly more bias. The large variability is because the size of the background window changes with spot size. To overcome this, a non-adaptive region-based method is implemented. Its bias and variability are comparable to that of the rank filter. CONCLUSION: The performance of more advanced rank filters is equal to the best region-based methods. However, in order to get unbiased estimates these filters have to be implemented with great care. The performance of morphological opening is in general poor with a substantial spatial-dependent bias

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