The rupture of thin-cap fibroatheroma accounts for most acute coronary events. Optical Coherence Tomography (OCT) allows quantification of fibrous cap (FC) thickness in vivo. Conventional manual analysis, by visually determining the thinnest part of the FC is subject to inter-observer variability and does not capture the 3-D morphology of the FC. We propose and validate a computer-aided method that allows volumetric analysis of FC. The radial FC boundary is semi-automatically segmented using a dynamic programming algorithm. The thickness at every point of the FC boundary, along with 3-D morphology of the FC, can be quantified. The method was validated against three experienced OCT image analysts in 14 lipid-rich lesions. The proposed method may advance our understanding of the mechanisms behind plaque rupture and improve disease management.
(100.0100) Image processing; (110.4500) Optical coherence tomography
Breast cancer is one of the leading causes of cancer death for women all over the world and mammography is thought of as one of the main tools for early detection of breast cancer. In order to detect the breast cancer, computer aided technology has been introduced. In computer aided cancer detection, the detection and segmentation of mass are very important. The shape of mass can be used as one of the factors to determine whether the mass is malignant or benign. However, many of the current methods are semi-automatic. In this paper, we investigate fully automatic segmentation method.
In this paper, a new mass segmentation algorithm is proposed. In the proposed algorithm, a fully automatic marker-controlled watershed transform is proposed to segment the mass region roughly, and then a level set is used to refine the segmentation. For over-segmentation caused by watershed, we also investigated different noise reduction technologies. Images from DDSM were used in the experiments and the results show that the new algorithm can improve the accuracy of mass segmentation.
The new algorithm combines the advantages of both methods. The combination of the watershed based segmentation and level set method can improve the efficiency of the segmentation. Besides, the introduction of noise reduction technologies can reduce over-segmentation.
The morphological properties of axons, such as their branching patterns and oriented structures, are of great interest for biologists in the study of the synaptic connectivity of neurons. In these studies, researchers use triple immunofluorescent confocal microscopy to record morphological changes of neuronal processes. Three-dimensional (3D) microscopy image analysis is then required to extract morphological features of the neuronal structures. In this article, we propose a highly automated 3D centerline extraction tool to assist in this task. For this project, the most difficult part is that some axons are overlapping such that the boundaries distinguishing them are barely visible. Our approach combines a 3D dynamic programming (DP) technique and marker-controlled watershed algorithm to solve this problem. The approach consists of tracking and updating along the navigation directions of multiple axons simultaneously. The experimental results show that the proposed method can rapidly and accurately extract multiple axon centerlines and can handle complicated axon structures such as cross-over sections and overlapping objects.
In previous research, a watershed-based algorithm was shown to be useful for automatic lesion segmentation in dermoscopy images, and was tested on a set of 100 benign and malignant melanoma images with the average of three sets of dermatologist-drawn borders used as the ground truth, resulting in an overall error of 15.98%. In this study, to reduce the border detection errors, a neural network classifier was utilized to improve the first-pass watershed segmentation; a novel “Edge Object Value (EOV) Threshold” method was used to remove large light blobs near the lesion boundary; and a noise removal procedure was applied to reduce the peninsula-shaped false-positive areas. As a result, an overall error of 11.09% was achieved.
Malignant Melanoma; Watershed; Image Processing; Segmentation; Neural Network
In this paper, we propose a method for automatic determination of position and orientation of spine in digitized spine X-rays using mathematical morphology. As the X-ray images are usually highly smeared, vertebrae segmentation is a complex process. The image is first coarsely segmented to obtain the location and orientation information of the spine. The state-of-the-art technique is based on the deformation model of a template, and as the vertebrae shape usually shows variation from case to case, accurate representation using a template is a difficult process. The proposed method makes use of the vertebrae morphometry and gray-scale profile of the spine. The top-hat transformation-based method is proposed to enhance the ridge points in the posterior boundary of the spine. For cases containing external objects such as ornaments, H-Maxima transform is used for segmentation and removal of these objects. The Radon transform is then used to estimate the location and orientation of the line joining the ridge point clusters appearing on the boundary of the vertebra body. The method was validated for 100 cervical spine X-ray images, and in all cases, the error in orientation was within the accepted tolerable limit of 15°. The average error was found to be 4.6°. A point on the posterior boundary was located with an accuracy of ±5.2 mm. The accurate information about location and orientation of thespine is necessary for fine-grained segmentation of the vertebrae using techniques such as active shape modeling. Accurate vertebrae segmentation is needed in successful feature extraction for applications such as content-based image retrieval of biomedical images.
Vertebrae segmentation; spine X-ray; content-based image retrieval; mathematical morphology
Extraction of the brain — i.e. cerebrum, cerebellum, and brain stem — from T1-weighted structural magnetic resonance images is an important initial step in neuroimage analysis. Although automatic algorithms are available, their inconsistent handling of the cortical mantle often requires manual interaction, thereby reducing their effectiveness. This paper presents a fully automated brain extraction algorithm that incorporates elastic registration, tissue segmentation, and morphological techniques which are combined by a watershed principle, while paying special attention to the preservation of the boundary between the gray matter and the cerebrospinal fluid. The approach was evaluated by comparison to a manual rater, and compared to several other leading algorithms on a publically available data set of brain images using the Dice coefficient and containment index as performance metrics. The qualitative and quantitative impact of this initial step on subsequent cortical surface generation is also presented. Our experiments demonstrate that our approach is quantitatively better than six other leading algorithms (with statistical significance on modern T1-weighted MR data). We also validated the robustness of the algorithm on a very large data set of over one thousand subjects, and showed that it can replace an experienced manual rater as preprocessing for a cortical surface extraction algorithm with statistically insignificant differences in cortical surface position.
Brain extraction; skull stripping; watershed principle; segmentation; medical image processing
Automatic lesion segmentation is an important part of computer-based image analysis of pigmented skin lesions. In this research, a watershed algorithm is developed and investigated for adequacy of skin lesion segmentation in dermoscopy images.
Hair, black border and vignette removal methods are introduced as preprocessing steps. The flooding variant of the watershed segmentation algorithm was implemented with novel features adapted to this domain. An outer bounding box, determined by a difference function derived from horizontal and vertical projection functions, is added to estimate the lesion area, and the lesion area error is reduced by a linear estimation function. As a post-processing step, a second-order B-Spline smoothing method is introduced to smooth the watershed border.
Using the average of three sets of dermatologist-drawn borders as the ground truth, an overall error of 15.98% was obtained using the watershed technique.
The implementation of the flooding variant of the watershed algorithm presented here allows satisfactory automatic segmentation of pigmented skin lesions.
malignant melanoma; watershed; image processing; segmentation
Two-dimensional echocardiography (2D-echo) allows the evaluation of cardiac structures and their movements. A wide range of clinical diagnoses are based on the performance of the left ventricle. The evaluation of myocardial function is typically performed by manual segmentation of the ventricular cavity in a series of dynamic images. This process is laborious and operator dependent. The automatic segmentation of the left ventricle in 4-chamber long-axis images during diastole is troublesome, because of the opening of the mitral valve.
This work presents a method for segmentation of the left ventricle in dynamic 2D-echo 4-chamber long-axis images over the complete cardiac cycle. The proposed algorithm is based on classic image processing techniques, including time-averaging and wavelet-based denoising, edge enhancement filtering, morphological operations, homotopy modification, and watershed segmentation. The proposed method is semi-automatic, requiring a single user intervention for identification of the position of the mitral valve in the first temporal frame of the video sequence. Image segmentation is performed on a set of dynamic 2D-echo images collected from an examination covering two consecutive cardiac cycles.
The proposed method is demonstrated and evaluated on twelve healthy volunteers. The results are quantitatively evaluated using four different metrics, in a comparison with contours manually segmented by a specialist, and with four alternative methods from the literature. The method's intra- and inter-operator variabilities are also evaluated.
The proposed method allows the automatic construction of the area variation curve of the left ventricle corresponding to a complete cardiac cycle. This may potentially be used for the identification of several clinical parameters, including the area variation fraction. This parameter could potentially be used for evaluating the global systolic function of the left ventricle.
Due to severe presence of speckle noise, poor image contrast and irregular lesion shape, it is challenging to build a fully automatic detection and classification system for breast ultrasonic images. In this paper, a novel and effective computer-aided method including generation of a region of interest (ROI), segmentation and classification of breast tumor is proposed without any manual intervention. By incorporating local features of texture and position, a ROI is firstly detected using a self-organizing map neural network. Then a modified Normalized Cut approach considering the weighted neighborhood gray values is proposed to partition the ROI into clusters and get the initial boundary. In addition, a regional-fitting active contour model is used to adjust the few inaccurate initial boundaries for the final segmentation. Finally, three textures and five morphologic features are extracted from each breast tumor; whereby a highly efficient Affinity Propagation clustering is used to fulfill the malignancy and benign classification for an existing database without any training process. The proposed system is validated by 132 cases (67 benignancies and 65 malignancies) with its performance compared to traditional methods such as level set segmentation, artificial neural network classifiers, and so forth. Experiment results show that the proposed system, which needs no training procedure or manual interference, performs best in detection and classification of ultrasonic breast tumors, while having the lowest computation complexity.
Breast ultrasonic images; fully automatic; region of interest; Normalized Cut; Affinity Propagation clustering.
In this report, a novel technique is proposed for computer-aided automatic extraction of microcalcifications in a digital mammogram. First, the microcalcifications are detected by morphological filtering, followed by entropy-based thresholding. Next, the microcalcifications are segmented by computing regional watershed. The proposed automatic technique is designed to serve as a visual aid to radiologists. Its efficacy is demonstrated through experimental results.
Cell boundary segmentation in live cell image sequences is the first step towards quantitative analysis of cell motion and behavior. The time lapse microscopy imaging produces large volumes of image sequence collections which requires fast and robust automatic segmentation of cell boundaries to utilize further automated tools such as cell tracking to quantify and classify cell behavior. This paper presents a methodology that is based on utilizing the temporal context of the cell image sequences to accurately delineate the boundaries of non-homogeneous cells. A novel flux tensor-based detection of moving cells provides initial localization that is further refined by a multi-feature level set-based method using an efficient additive operator splitting scheme. The segmentation result is processed by a watershed-based algorithm to avoid merging boundaries of neighboring cells. By utilizing robust features, the level-set algorithm produces accurate segmentation for non-homogeneous cells with concave shapes and varying intensities.
Automatic image analysis of histopathology specimens would help the early detection of blood cancer. The first step for automatic image analysis is segmentation. However, touching cells bring the difficulty for traditional segmentation algorithms. In this paper, we propose a novel algorithm which can reliably handle touching cells segmentation. Robust estimation and color active contour models are used to delineate the outer boundary. Concave points on the boundary and inner edges are automatically detected. A concave vertex graph is constructed from these points and edges. By minimizing a cost function based on morphological characteristics, we recursively calculate the optimal path in the graph to separate the touching cells. The algorithm is computationally efficient and has been tested on two large clinical dataset which contain 207 images and 3898 images respectively. Our algorithm provides better results than other studies reported in the recent literature.
Snake or active contours are extensively used in computer vision and medical image processing applications, and particularly to locate object boundaries. Yet problems associated with initialization and the poor convergence to boundary concavities have limited their utility. The new method of external force for active contours, which is called gradient vector flow (GVF), was recently introduced to address the problems.
This paper presents an automatic initialization value of the snake algorithm for the segmentation of the chest wall. Snake algorithms are required to have manually drawn initial contours, so this needs automatic initialization. In this paper, our proposed algorithm is the mean shape for automatic initialization in the GVF.
The GVF is calculated as a diffusion of the gradient vectors of a gray-level or binary edge map derived from the medical images. Finally, the mean shape coordinates are used to automatic initialize thepoint of the snake. The proposed algorithm is composed of three phases: the landmark phase, the procrustes shape distance metric phase and aligning a set of shapes phase. The experiments showed the good performance of our algorithm in segmenting the chest wall by chest computed tomography.
An error analysis for the active contours results on simulated test medical images is also presented. We showed that GVF has a large capture range and it is able to move a snake into boundary concavities. Therefore, the suggested algorithm is better than the traditional potential forces of image segmentation.
Active Contour Model; Automatic Initialization; Mean Shape; Gradient Vector Flow; Computed Tomography
Correlation of information from multiple-view mammograms (e.g., MLO and CC views, bilateral views, or current and prior mammograms) can improve the performance of breast cancer diagnosis by radiologists or by computer. The nipple is a reliable and stable landmark on mammograms for the registration of multiple mammograms. However, accurate identification of nipple location on mammograms is challenging because of the variations in image quality and in the nipple projections, resulting in some nipples being nearly invisible on the mammograms. In this study, we developed a computerized method to automatically identify the nipple location on digitized mammograms. First, the breast boundary was obtained using a gradient-based boundary tracking algorithm, and then the gray level profiles along the inside and outside of the boundary were identified. A geometric convergence analysis was used to limit the nipple search to a region of the breast boundary. A two-stage nipple detection method was developed to identify the nipple location using the gray level information around the nipple, the geometric characteristics of nipple shapes, and the texture features of glandular tissue or ducts which converge toward the nipple. At the first stage, a rule-based method was designed to identify the nipple location by detecting significant changes of intensity along the gray level profiles inside and outside the breast boundary and the changes in the boundary direction. At the second stage, a texture orientation-field analysis was developed to estimate the nipple location based on the convergence of the texture pattern of glandular tissue or ducts towards the nipple. The nipple location was finally determined from the detected nipple candidates by a rule-based confidence analysis. In this study, 377 and 367 randomly selected digitized mammograms were used for training and testing the nipple detection algorithm, respectively. Two experienced radiologists identified the nipple locations which were used as the gold standard. In the training data set, 301 nipples were positively identified and were referred to as visible nipples. Seventy six nipples could not be positively identified and were referred to as invisible nipples. The radiologists provided their estimation of the nipple locations in the latter group for comparison with the computer estimates. The computerized method could detect 89.37% (269/301) of the visible nipples and 69.74% (53/76) of the invisible nipples within 1 cm of the gold standard. In the test data set, 298 and 69 of the nipples were classified as visible and invisible, respectively. 92.28% (275/298) of the visible nipples and 53.62% (37/69) of the invisible nipples were identified within 1 cm of the gold standard. The results demonstrate that the nipple locations on digitized mammograms can be accurately detected if they are visible and can be reasonably estimated if they are invisible. Automated nipple detection will be an important step towards multiple image analysis for CAD.
computer-aided detection; mammography; nipple detection; texture orientation field analysis
Automated segmentation of time-lapse images is a method to facilitate the understanding of the intricate biological progression, e.g., cancer cell migration. To address this problem, we introduce a shape representation enhancement over popular snake models in the context of confident scale-space such that a higher level of interpretation can hopefully be achieved. Our proposed system consists of a hierarchical analytic framework including feedback loops, self-adaptive and demand-adaptive adjustment, incorporating a steerable boundary detail term constraint based on multiscale B-spline interpolation. To minimize the noise interference inherited from microscopy acquisition, the coarse boundary derived from the initial segmentation with refined watershed line is coupled with microscopy compensation using the mean shift filtering. A progressive approximation is applied to achieve represented as a balance between a relief function of watershed algorithm and local minima concerning multi-scale optimality, convergence, and robust constraints. Experimental results show that the proposed method overcomes problems with spurious branches, arbitrary gaps, low contrast boundaries and low signal-to-noise ratio. The proposed system has the potential to serve as an automated data processing tool for cell migration applications.
cellular image segmentation; 3T3 cell; time-lapse microscopy; snake model; multiscale detail detection; mean shift filtering
We are presenting a program for interactive segmentation of tomographic maps, based on objective criteria so as to yield reproducible results. The strategy starts with the automatic segmentation of the entire volume with the watershed algorithm in 3D. The watershed regions are clustered successively by supervised classification, allowing the segmentation of known organelles, such as membranes, vesicles and microtubules. These organelles are processed with topological models and input parameters manually derived from the tomograms. After known organelles are extracted from the volume, all other watershed regions can be organized into homogeneous assemblies on the basis of their densities. To complete the process, all voxels in the volume are assigned either to the background or individual structures, which can then be extracted for visualization with any rendering technique.
The user interface of the program is written in Java, and computational routines are written in C. For some operations, involving the visualization of the tomogram, we refer to existing software, either open or commercial. While the program runs, a history file is created, that allows all parameters and other data to be saved for the purposes of comparison or exchange. Initially, the program was developed for the segmentation of synapses, and organelles belonging to these structures have thus far been the principal targets modeled with JUST. Since each organelle is clustered independently from the rest of the volume, however, the program can accommodate new models of different organelles as well as tomograms of other types of preparations of tissue, such as citoskeletal components in vitreous ice.
Electron tomography; Volume segmentation; Image processing; Feature extraction; Synapsis
Optical microscopy is becoming an important technique in drug discovery and life science research. The approaches used to analyze optical microscopy images are generally classified into two categories: automatic and manual approaches. However, the existing automatic systems are rather limited in dealing with large volume of time-lapse microscopy images because of the complexity of cell behaviors and morphological variance. On the other hand, manual approaches are very time-consuming. In this paper, we propose an effective automated, quantitative analysis system that can be used to segment, track, and quantize cell cycle behaviors of a large population of cells nuclei effectively and efficiently. We use adaptive thresholding and watershed algorithm for cell nuclei segmentation followed by a fragment merging method that combines two scoring models based on trend and no trend features. Using the context information of time-lapse data, the phases of cell nuclei are identified accurately via a Markov model. Experimental results show that the proposed system is effective for nuclei segmentation and phase identification.
Cell phase identification; continuous Markov model; nuclei segmentation; time-lapse fluorescence microscopy; tracking
Starches are the main storage polysaccharides in plants and are distributed widely throughout plants including seeds, roots, tubers, leaves, stems and so on. Currently, microscopic observation is one of the most important ways to investigate and analyze the structure of starches. The position, shape, and size of the starch granules are the main measurements for quantitative analysis. In order to obtain these measurements, segmentation of starch granules from the background is very important. However, automatic segmentation of starch granules is still a challenging task because of the limitation of imaging condition and the complex scenarios of overlapping granules.
We propose a novel method to segment starch granules in microscopic images. In the proposed method, we first separate starch granules from background using automatic thresholding and then roughly segment the image using watershed algorithm. In order to reduce the oversegmentation in watershed algorithm, we use the roundness of each segment, and analyze the gradient vector field to find the critical points so as to identify oversegments. After oversegments are found, we extract the features, such as the position and intensity of the oversegments, and use fuzzy c-means clustering to merge the oversegments to the objects with similar features. Experimental results demonstrate that the proposed method can alleviate oversegmentation of watershed segmentation algorithm successfully.
We present a new scheme for starch granules segmentation. The proposed scheme aims to alleviate the oversegmentation in watershed algorithm. We use the shape information and critical points of gradient vector flow (GVF) of starch granules to identify oversegments, and use fuzzy c-mean clustering based on prior knowledge to merge these oversegments to the objects. Experimental results on twenty microscopic starch images demonstrate the effectiveness of the proposed scheme.
Automatic aorta segmentation in thoracic computed tomography (CT) scans is important for aortic calcification quantification and to guide the segmentation of other central vessels. We propose an aorta segmentation algorithm consisting of an initial boundary detection step followed by 3D level set segmentation for refinement. Our algorithm exploits aortic cross-sectional circularity: we first detect aorta boundaries with a circular Hough transform on axial slices to detect ascending and descending aorta regions, and we apply the Hough transform on oblique slices to detect the aortic arch. The centers and radii of circles detected by Hough transform are fitted to smooth cubic spline functions using least-squares fitting. From these center and radius spline functions, we reconstruct an initial aorta surface using the Frenet frame. This reconstructed tubular surface is further refined with 3D level set evolutions. The level set framework we employ optimizes a functional that depends on both edge strength and smoothness terms and evolves the surface to the position of nearby edge location corresponding to the aorta wall. After aorta segmentation, we first detect the aortic calcifications with thresholding applied to the segmented aorta region. We then filter out the false positive regions due to nearby high intensity structures. We tested the algorithm on 45 CT scans and obtained a closest point mean error of 0.52 ± 0.10 mm between the manually and automatically segmented surfaces. The true positive detection rate of calcification algorithm was 0.96 over all CT scans.
Delineation of radiofrequency-ablation-induced coagulation (thermal lesion) boundaries is an important clinical problem that is not well addressed by conventional imaging modalities. Elastography, which produces images of the local strain after small, externally applied compressions, can be used for visualization of thermal coagulations. This paper presents an automated segmentation approach for thermal coagulations on 3-D elastographic data to obtain both area and volume information rapidly. The approach consists of a coarse-to-fine method for active contour initialization and a gradient vector flow, active contour model for deformable contour optimization with the help of prior knowledge of the geometry of general thermal coagulations. The performance of the algorithm has been shown to be comparable to manual delineation of coagulations on elastograms by medical physicists (r = 0.99 for volumes of 36 radiofrequency-induced coagulations). Furthermore, the automatic algorithm applied to elastograms yielded results that agreed with manual delineation of coagulations on pathology images (r = 0.96 for the same 36 lesions). This algorithm has also been successfully applied on in vivo elastograms.
Ablation; Active contour models; Breast tumor; Elasticity imaging; Elastography; Gradient vector flow; Image segmentation; Multiresolution; Snakes; Strain; 3-D ultrasound
Automated cell imaging systems facilitate fast and reliable analysis of biological events at the cellular level. In these systems, the first step is usually cell segmentation that greatly affects the success of the subsequent system steps. On the other hand, similar to other image segmentation problems, cell segmentation is an ill-posed problem that typically necessitates the use of domain-specific knowledge to obtain successful segmentations even by human subjects. The approaches that can incorporate this knowledge into their segmentation algorithms have potential to greatly improve segmentation results. In this work, we propose a new approach for the effective segmentation of live cells from phase contrast microscopy. This approach introduces a new set of “smart markers” for a marker-controlled watershed algorithm, for which the identification of its markers is critical. The proposed approach relies on using domain-specific knowledge, in the form of visual characteristics of the cells, to define the markers. We evaluate our approach on a total of 1,954 cells. The experimental results demonstrate that this approach, which uses the proposed definition of smart markers, is quite effective in identifying better markers compared to its counterparts. This will, in turn, be effective in improving the segmentation performance of a marker-controlled watershed algorithm.
We propose to segment two-dimensional CT scans traumatic
brain injuries with various methods. These methods are
hybrid, feature extraction, level sets, region growing, and
watershed which are analysed based upon their parametric
and nonparametric arguments. The pixel intensities, gradient
magnitude, affinity map, and catchment basins of these
methods are validated based upon various constraints evaluations.
In this article, we also develop a new methodology for
a computational pipeline that uses bilateral filtering, diffusion
properties, watershed, and filtering with mathematical
morphology operators for the contour extraction of the lesion
in the feature available based mainly on the gradient
function. The evaluations of the classification of these lesions
are very briefly outlined in this context and are being
undertaken by pattern recognition in another paper work.
High content neuron image processing is considered as an important method for quantitative neurobiological studies. The main goal of analysis in this paper is to provide automatic image processing approaches to process neuron images for studying neuron mechanism in high content screening. In the nuclei channel, all nuclei are segmented and detected by applying the gradient vector field based watershed. Then the neuronal nuclei are selected based on the soma region detected in neurite channel. In neurite images, we propose a novel neurite centerline extraction approach using the improved line-pixel detection technique. The proposed neurite tracing method can detect the curvilinear structure more accurately compared with the current existing methods. An interface called NeuriteIQ based on the proposed algorithms is developed finally for better application in high content screening.
High content screening; Microscopy image; Nuclei segmentation; Neurite outgrowth; Line-pixel detection; Branch area
In this paper, an unsupervised algorithm, called the Independent Histogram Pursuit (IHP), for segmenting dermatological lesions is proposed. The algorithm estimates a set of linear combinations of image bands that enhance different structures embedded in the image. In particular, the first estimated combination enhances the contrast of the lesion to facilitate its segmentation. Given an N-band image, this first combination corresponds to a line in N dimensions, such that the separation between the two main modes of the histogram obtained by projecting the pixels onto this line, is maximized. The remaining combinations are estimated in a similar way under the constraint of being orthogonal to those already computed. The performance of the algorithm is tested on five different dermatological datasets. The results obtained on these datasets indicate the robustness of the algorithm and its suitability to deal with different types of dermatological lesions. The boundary detection precision using k-means segmentation was close to 97%. The proposed algorithm can be easily combined with the majority of classification algorithms.
Boundary detection; classification; dermoscopy; exploratory data analysis; feature extraction; genetic algorithms; independent component analysis; projection pursuit
The introduction of fast digital slide scanners that provide whole slide images has led to a revival of interest in image analysis applications in pathology. Segmentation of cells and nuclei is an important first step towards automatic analysis of digitized microscopy images. We therefore developed an automated nuclei segmentation method that works with hematoxylin and eosin (H&E) stained breast cancer histopathology images, which represent regions of whole digital slides. The procedure can be divided into four main steps: 1) pre-processing with color unmixing and morphological operators, 2) marker-controlled watershed segmentation at multiple scales and with different markers, 3) post-processing for rejection of false regions and 4) merging of the results from multiple scales. The procedure was developed on a set of 21 breast cancer cases (subset A) and tested on a separate validation set of 18 cases (subset B). The evaluation was done in terms of both detection accuracy (sensitivity and positive predictive value) and segmentation accuracy (Dice coefficient). The mean estimated sensitivity for subset A was 0.875 (±0.092) and for subset B 0.853 (±0.077). The mean estimated positive predictive value was 0.904 (±0.075) and 0.886 (±0.069) for subsets A and B, respectively. For both subsets, the distribution of the Dice coefficients had a high peak around 0.9, with the vast majority of segmentations having values larger than 0.8.