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Background Cryo-electron tomography is an important and powerful technique to explore the structure, abundance, and location of ultrastructure in a near-native state. It contains detailed information of all macromolecular complexes in a sample cell. However, due to the compact and crowded status, the missing edge effect, and low signal to noise ratio (SNR), it is extremely challenging to recover such information with existing image processing methods. Cryo-electron tomogram simulation is an effective solution to test and optimize the performance of the above image processing methods. The simulated images could be regarded as the labeled data which covers a wide range of macromolecular complexes and ultrastructure. To approximate the crowded cellular environment, it is very important to pack these heterogeneous structures as tightly as possible. Besides, simulating non-deformable and deformable components under a unified framework also need to be achieved. Result In this paper, we proposed a unified framework for simulating crowded cryo-electron tomogram images including non-deformable macromolecular complexes and deformable ultrastructures. A macromolecule was approximated using multiple balls with fixed relative positions to reduce the vacuum volume. A ultrastructure, such as membrane and filament, was approximated using multiple balls with flexible relative positions so that this structure could deform under force field. In the experiment, 400 macromolecules of 20 representative types were packed into simulated cytoplasm by our framework, and numerical verification proved that our method has a smaller volume and higher compression ratio than the baseline single-ball model. We also packed filaments, membranes and macromolecules together, to obtain a simulated cryo-electron tomogram image with deformable structures. The simulated results are closer to the real Cryo-ET, making the analysis more difficult. The DOG particle picking method and the image segmentation method are tested on our simulation data, and the experimental results show that these methods still have much room for improvement. Conclusion The proposed multi-ball model can achieve more crowded packaging results and contains richer elements with different properties to obtain more realistic cryo-electron tomogram simulation. This enables users to simulate cryo-electron tomogram images with non-deformable macromolecular complexes and deformable ultrastructures under a unified framework. To illustrate the advantages of our framework in improving the compression ratio, we calculated the volume of simulated macromolecular under our multi-ball method and traditional single-ball method. We also performed the packing experiment of filaments and membranes to demonstrate the simulation ability of deformable structures. Our method can be used to do a benchmark by generating large labeled cryo-ET dataset and evaluating existing image processing methods. Since the content of the simulated cryo-ET is more complex and crowded compared with previous ones, it will pose a greater challenge to existing image processing methods.

Background Immunofluorescent confocal microscopy uses labeled antibodies as probes against specific macromolecules to discriminate between multiple cell types. For images of the developmental mouse lung, these cells are themselves organized into densely packed higher-level anatomical structures. These types of images can be challenging to segment automatically for several reasons, including the relevance of biomedical context, dependence on the specific set of probes used, prohibitive cost of generating labeled training data, as well as the complexity and dense packing of anatomical structures in the image. The use of an application ontology helps surmount these challenges by combining image data with its metadata to provide a meaningful biological context, modeled after how a human expert would make use of contextual information to identify histological structures, that constrains and simplifies the process of segmentation and object identification. Results We propose an innovative approach for the semi-supervised analysis of complex and densely packed anatomical structures from immunofluorescent images that utilizes an application ontology to provide a simplified context for image segmentation and object identification. We describe how the logical organization of biological facts in the form of an ontology can provide useful constraints that facilitate automatic processing of complex images. We demonstrate the results of ontology-guided segmentation and object identification in mouse developmental lung images from the Bioinformatics REsource ATlas for the Healthy lung database of the Molecular Atlas of Lung Development (LungMAP1) program Conclusion We describe a novel ontology-guided approach to segmentation and classification of complex immunofluorescence images of the developing mouse lung. The ontology is used to automatically generate constraints for each image based on its biomedical context, which facilitates image segmentation and classification.

A common gynecological disease in the world is breast cancer that early diagnosis of this disease can be very effective in its treatment. The use of image processing methods and pattern recognition techniques in automatic breast detection from mammographic images decreases human errors and increments the rapidity of diagnosis. In this paper, mammographic images are analyzed using image processing techniques and a pipeline structure for the diagnosis of the cancerous masses. In the first stage, the quality of mammogram images and the contrast of abnormal areas in the image are improved by using image contrast improvement and a noise decline. A method based on color space is then used for image segmentation that is followed by mathematical morphology. Then, for feature image extraction, a combined gray-level cooccurrence matrix (GLCM) and discrete wavelet transform (DWT) method is used. At last, a new optimized version of convolutional neural network (CNN) and a new improved metaheuristic, called Advanced Thermal Exchange Optimizer, are used for the classification of the features. A comparison of the simulations of the proposed technique with three different techniques from the literature applied on the MIAS mammogram database is performed to show its superiority. Results show that the accuracy of diagnosing cancer cases for the proposed method and applied on the MIAS database is 93.79%, and sensitivity and specificity are obtained 96.89% and 67.7%, respectively.

International audience; Traditional methods to measure spatio-temporal variations in biomass rely on a labor-intensive destructive sampling of the crop. In this paper, we present a high-throughput phenotyping approach for the estimation of Above-Ground Biomass Dynamics (AGBD) using an unmanned aerial system. Multispectral imagery was acquired and processed by using the proposed segmentation method called GFKuts, that optimally labels the plot canopy based on a Gaussian mixture model, a Montecarlo based K-means, and a guided image filtering. Accurate plot segmentation results enabled the extraction of several canopy features associated with biomass yield. Machine learning algorithms were trained to estimate the AGBD according to the growth stages of the crop and the physiological response of two rice genotypes under lowland and upland production systems. Results report AGBD estimation correlations with an average of r = 0.95 and R 2 = 0.91 according to the experimental data. We compared our segmentation method against a traditional technique based on clustering. A comprehensive improvement of 13% in the biomass correlation was obtained thanks to the segmentation method proposed herein.

Image segmentation, as a key step of microarray image processing, is crucial for obtaining the spot expressions simultaneously. However, state-of-art clustering-based segmentation algorithms are sensitive to noises. To solve this problem and improve the segmentation accuracy, in this article, several improvements are introduced into the fast and simple clustering methods (K-means and Fuzzy C means). Firstly, a contrast enhancement algorithm is implemented in image preprocessing to improve the gridding precision. Secondly, the data-driven means are proposed for cluster center initialization, instead of usual random setting. The third improvement is that the multi features, including intensity features, spatial features, and shape features, are implemented in feature selection to replace the sole pixel intensity feature used in the traditional clustering-based methods to avoid taking noises as spot pixels. Moreover, the principal component analysis is adopted for various feature extraction. Finally, an adaptive adjustment algorithm is proposed based on data mining and learning for further dealing with the missing spots or low contrast spots. Experiments on real and simulation data sets indicate that the proposed improvements made our proposed method obtains higher segmented precision than the traditional K-means and Fuzzy C means clustering methods.

With the increase of the camera resolution, the number of pixels contained in froth image is increased, which brings many challenges to image segmentation. Froth size and distribution are the important index in froth flotation. The segmentation of froth images is always a problem in building flotation model. In segmenting froth images, Otsu method is usually used to get a binary image for classification of froth images, this method can get a satisfactory segmentation result. However, each gray level is required to calculate each of the between-class variance, it takes a longer time in froth images with a large number of pixels. To solve this problem, an improved method is proposed in this paper. Most froth images have the pixel distribution characteristic that the gray histogram curve is a sawtooth shape. The proposed method uses polynomial to fit the curve of gray histogram and takes the characteristic of gray histogram's valley into consideration in Otsu method. Two performance comparison methods are introduced and used. Experimental comparison between Otsu method and the proposed method shows that the proposed method has a satisfactory image segmentation with a low computing time.

Background The protein ki67 (pki67) is a marker of tumor aggressiveness, and its expression has been proven to be useful in the prognostic and predictive evaluation of several types of tumors. To numerically quantify the pki67 presence in cancerous tissue areas, pathologists generally analyze histochemical images to count the number of tumor nuclei marked for pki67. This allows estimating the ki67-index, that is the percentage of tumor nuclei positive for pki67 over all the tumor nuclei. Given the high image resolution and dimensions, its estimation by expert clinicians is particularly laborious and time consuming. Though automatic cell counting techniques have been presented so far, the problem is still open. Results In this paper we present a novel automatic approach for the estimations of the ki67-index. The method starts by exploiting the STRESS algorithm to produce a color enhanced image where all pixels belonging to nuclei are easily identified by thresholding, and then separated into positive (i.e. pixels belonging to nuclei marked for pki67) and negative by a binary classification tree. Next, positive and negative nuclei pixels are processed separately by two multiscale procedures identifying isolated nuclei and separating adjoining nuclei. The multiscale procedures exploit two Bayesian classification trees to recognize positive and negative nuclei-shaped regions. Conclusions The evaluation of the computed results, both through experts’ visual assessments and through the comparison of the computed indexes with those of experts, proved that the prototype is promising, so that experts believe in its potential as a tool to be exploited in the clinical practice as a valid aid for clinicians estimating the ki67-index. The MATLAB source code is open source for research purposes.

Assessing the cytoplasmic uptake of fluorescently-tagged drugs in heterogeneous cell types currently involves time-consuming manual segmentation of confocal microscopy images. We developed a set of methods that incorporate map algebra techniques to facilitate and expedite image segmentation, particularly of the parenchyma of intermediate cells in the stria vascularis of the inner ear. Map algebra is used to apply a convolution kernel to pixel neighborhoods to create a masking image to select pixels in the original image for further operations. Here, we describe the utility of integrated intensity-based, percentile-based, and local autocorrelation-based methods to automate segmentation of images into putative morphological regions for pixel intensity analysis. Integrated intensity-based methods are variants of watershed segmentation tools that determine morphological boundaries from rates of change in integrated pixel intensity. Percentile- and local autocorrelation-based methods evolved out of the process of developing map algebra- and integrated intensity-based tools. We identified several simplifications that are surprisingly effective for image segmentation and pixel intensity analysis. These methods were empirically validated on three levels: first, the algorithms were developed based on iterations of inspected results; second, algorithms were tested for various types of robustness; and third, developed algorithms were validated against results from manually-segmented images. We conclude the key to automated segmentation is supervision of output data.

In this paper, we propose a novel algorithm for medical image segmentation, which combines the density peaks clustering (DPC) with the fruit fly optimization algorithm, and it has the following advantages. Firstly, it avoids the problem of DPC that needs to artificially select parameters (such as the number of clusters) in its decision graph and thus can automatically determine their values. Secondly, our algorithm uses random step size, instead of the fixed step size as in the fruit fly optimization algorithm, which helps avoid falling into local optima. Thirdly, our algorithm selects the cut-off distance and the cluster centers using the image entropy value and can better capture the structures of the image. Experiments on benchmark dataset and proprietary dataset show that our algorithm can adaptively segment medical images with faster convergence and better robustness.

We present a method combining affinity prediction with region agglomeration, which improves significantly upon the state of the art of neuron segmentation from electron microscopy (EM) in accuracy and scalability. Our method consists of a 3D U-Net, trained to predict affinities between voxels, followed by iterative region agglomeration. We train using a structured loss based on Malis, encouraging topologically correct segmentations obtained from affinity thresholding. Our extension consists of two parts: First, we present a quasi-linear method to compute the loss gradient, improving over the original quadratic algorithm. Second, we compute the gradient in two separate passes to avoid spurious gradient contributions in early training stages. Our predictions are accurate enough that simple learning-free percentile-based agglomeration outperforms more involved methods used earlier on inferior predictions. We present results on three diverse EM datasets, achieving relative improvements over previous results of 27, 15, and 250 percent. Our findings suggest that a single method can be applied to both nearly isotropic block-face EM data and anisotropic serial sectioned EM data. The runtime of our method scales linearly with the size of the volume and achieves a throughput of $\sim$∼ 2.6 seconds per megavoxel, qualifying our method for the processing of very large datasets.

In general, a single thresholding technique is developed or enhanced to separate foreground objects from background for a domain of images. This idea may not generate satisfactory results for all images in a dataset, since different images may require different types of thresholding methods for proper binarization or segmentation. To overcome this limitation, in this study, we propose a novel approach called “super-thresholding” that utilizes a supervised classifier to decide an appropriate thresholding method for a specific image. This method provides a generic framework that allows selection of the best thresholding method among different thresholding techniques that are beneficial for the problem domain. A classifier model is built using features extracted priori from the original image only or posteriori by analyzing the outputs of thresholding methods and the original image. This model is applied to identify the thresholding method for new images of the domain. We performed our method on protein crystallization images, and then we compared our results with six thresholding techniques. Numerical results are provided using four different correctness measurements. Super-thresholding outperforms the best single thresholding method around 10 percent, and it gives the best performance for protein crystallization dataset in our experiments.

Early brain tumor detection and diagnosis are critical to clinics. Thus segmentation of focused tumor area needs to be accurate, efficient, and robust. In this paper, we propose an automatic brain tumor segmentation method based on Convolutional Neural Networks (CNNs). Traditional CNNs focus only on local features and ignore global region features, which are both important for pixel classification and recognition. Besides, brain tumor can appear in any place of the brain and be any size and shape in patients. We design a three-stream framework named as multiscale CNNs which could automatically detect the optimum top-three scales of the image sizes and combine information from different scales of the regions around that pixel. Datasets provided by Multimodal Brain Tumor Image Segmentation Benchmark (BRATS) organized by MICCAI 2013 are utilized for both training and testing. The designed multiscale CNNs framework also combines multimodal features from T1, T1-enhanced, T2, and FLAIR MRI images. By comparison with traditional CNNs and the best two methods in BRATS 2012 and 2013, our framework shows advances in brain tumor segmentation accuracy and robustness.

A novel method for finger vein pattern extraction from infrared images is presented. This method involves four steps: preprocessing which performs local normalization of the image intensity, image enhancement, image segmentation, and finally postprocessing for image cleaning. In the image enhancement step, an image which will be both smooth and similar to the original is sought. The enhanced image is obtained by minimizing the objective function of a modified separable Mumford Shah Model. Since, this minimization procedure is computationally intensive for large images, a local application of the Mumford Shah Model in small window neighborhoods is proposed. The finger veins are located in concave nonsmooth regions and, so, in order to distinct them from the other tissue parts, all the differences between the smooth neighborhoods, obtained by the local application of the model, and the corresponding windows of the original image are added. After that, veins in the enhanced image have been sufficiently emphasized. Thus, after image enhancement, an accurate segmentation can be obtained readily by a local entropy thresholding method. Finally, the resulted binary image may suffer from some misclassifications and, so, a postprocessing step is performed in order to extract a robust finger vein pattern.

International audience; Connected operators provide well-established solutions for digital image processing, typically in conjunction with hierarchical schemes. In graph-based frameworks, such operators basically rely on symmetric adjacency relations between pixels. In this article, we introduce a notion of directed connected operators for hierarchical image processing, by also considering non-symmetric adjacency relations. The induced image representation models are no longer partition hierarchies (i.e., trees), but directed acyclic graphs that generalize standard morphological tree structures such as component trees, binary partition trees or hierarchical watersheds. We describe how to efficiently build and handle these richer data structures, and we illustrate the versatility of the proposed framework in image filtering and image segmentation.

Image segmentation plays an important role in medical image processing. Fuzzyc-means (FCM) clustering is one of the popular clustering algorithms for medical image segmentation. However, FCM has the problems of depending on initial clustering centers, falling into local optimal solution easily, and sensitivity to noise disturbance. To solve these problems, this paper proposes a hybrid artificial fish swarm algorithm (HAFSA). The proposed algorithm combines artificial fish swarm algorithm (AFSA) with FCM whose advantages of global optimization searching and parallel computing ability of AFSA are utilized to find a superior result. Meanwhile, Metropolis criterion and noise reduction mechanism are introduced to AFSA for enhancing the convergence rate and antinoise ability. The artificial grid graph and Magnetic Resonance Imaging (MRI) are used in the experiments, and the experimental results show that the proposed algorithm has stronger antinoise ability and higher precision. A number of evaluation indicators also demonstrate that the effect of HAFSA is more excellent than FCM and suppressed FCM (SFCM).