We present a conceptually simple framework for object instance segmentation called Contour Proposal Network (CPN), which detects possibly overlapping objects in an image while simultaneously fitting closed object contours using an interpretable, fixed-sized representation based on Fourier Descriptors. The CPN can incorporate state of the art object detection architectures as backbone networks into a single-stage instance segmentation model that can be trained end-to-end. We construct CPN models with different backbone networks, and apply them to instance segmentation of cells in datasets from different modalities. In our experiments, we show CPNs that outperform U-Nets and Mask R-CNNs in instance segmentation accuracy, and present variants with execution times suitable for real-time applications. The trained models generalize well across different domains of cell types. Since the main assumption of the framework are closed object contours, it is applicable to a wide range of detection problems also outside the biomedical domain. An implementation of the model architecture in PyTorch is freely available.
Cervical cancer, one of the most common fatal cancers among women, can be prevented by regular screening to detect any precancerous lesions at early stages and treat them. Pap smear test is a widely performed screening technique for early detection of cervical cancer, whereas this manual screening method suffers from high false-positive results because of human errors. To improve the manual screening practice, machine learning (ML) and deep learning (DL) based computer-aided diagnostic (CAD) systems have been investigated widely to classify cervical pap cells. Most of the existing researches require pre-segmented images to obtain good classification results, whereas accurate cervical cell segmentation is challenging because of cell clustering. Some studies rely on handcrafted features, which cannot guarantee the classification stage's optimality. Moreover, DL provides poor performance for a multiclass classification task when there is an uneven distribution of data, which is prevalent in the cervical cell dataset. This investigation has addressed those limitations by proposing DeepCervix, a hybrid deep feature fusion (HDFF) technique based on DL to classify the cervical cells accurately. Our proposed method uses various DL models to capture more potential information to enhance classification performance. Our proposed HDFF method is tested on the publicly available SIPAKMED dataset and compared the performance with base DL models and the LF method. For the SIPAKMED dataset, we have obtained the state-of-the-art classification accuracy of 99.85%, 99.38%, and 99.14% for 2-class, 3-class, and 5-class classification. Moreover, our method is tested on the Herlev dataset and achieves an accuracy of 98.32% for binary class and 90.32% for 7-class classification.
Cell segmentation and tracking in microscopy images are of great significance to new discoveries in biology and medicine. In this study, we propose a novel approach to combine cell segmentation and cell tracking into a unified end-to-end deep learning based framework, where cell detection and segmentation are performed with a current instance segmentation pipeline and cell tracking is implemented by integrating Siamese Network with the pipeline. Besides, tracking performance is improved by incorporating spatial information into the network and fusing spatial and visual prediction. Our approach was evaluated on the DeepCell benchmark dataset. Despite being simple and efficient, our method outperforms state-of-the-art algorithms in terms of both cell segmentation and cell tracking accuracies.
We present AURA-net, a convolutional neural network (CNN) for the segmentation of phase-contrast microscopy images. AURA-net uses transfer learning to accelerate training and Attention mechanisms to help the network focus on relevant image features. In this way, it can be trained efficiently with a very limited amount of annotations. Our network can thus be used to automate the segmentation of datasets that are generally considered too small for deep learning techniques. AURA-net also uses a loss inspired by active contours that is well-adapted to the specificity of phase-contrast images, further improving performance. We show that AURA-net outperforms state-of-the-art alternatives in several small (less than 100images) datasets.
Automatic cell segmentation is an essential step in the pipeline of computer-aided diagnosis (CAD), such as the detection and grading of breast cancer. Accurate segmentation of cells can not only assist the pathologists to make a more precise diagnosis, but also save much time and labor. However, this task suffers from stain variation, cell inhomogeneous intensities, background clutters and cells from different tissues. To address these issues, we propose an Attention Enforced Network (AENet), which is built on spatial attention module and channel attention module, to integrate local features with global dependencies and weight effective channels adaptively. Besides, we introduce a feature fusion branch to bridge high-level and low-level features. Finally, the marker controlled watershed algorithm is applied to post-process the predicted segmentation maps for reducing the fragmented regions. In the test stage, we present an individual color normalization method to deal with the stain variation problem. We evaluate this model on the MoNuSeg dataset. The quantitative comparisons against several prior methods demonstrate the superiority of our approach.
Single cell segmentation is critical and challenging in live cell imaging data analysis. Traditional image processing methods and tools require time-consuming and labor-intensive efforts of manually fine-tuning parameters. Slight variations of image setting may lead to poor segmentation results. Recent development of deep convolutional neural networks(CNN) provides a potentially efficient, general and robust method for segmentation. Most existing CNN-based methods treat segmentation as a pixel-wise classification problem. However, three unique problems of cell images adversely affect segmentation accuracy: lack of established training dataset, few pixels on cell boundaries, and ubiquitous blurry features. The problem becomes especially severe with densely packed cells, where a pixel-wise classification method tends to identify two neighboring cells with blurry shared boundary as one cell, leading to poor cell count accuracy and affecting subsequent analysis. Here we developed a different learning strategy that combines strengths of CNN and watershed algorithm. The method first trains a CNN to learn Euclidean distance transform of binary masks corresponding to the input images. Then another CNN is trained to detect individual cells in the Euclidean distance transform. In the third step, the watershed algorithm takes the outputs from the previous steps as inputs and performs the segmentation. We tested the combined method and various forms of the pixel-wise classification algorithm on segmenting fluorescence and transmitted light images. The new method achieves similar pixel accuracy but significant higher cell count accuracy than pixel-wise classification methods do, and the advantage is most obvious when applying on noisy images of densely packed cells.
Automated red blood cell (RBC) classification on blood smear images helps hematologists to analyze RBC lab results in a reduced time and cost. However, overlapping cells can cause incorrect predicted results, and so they have to be separated into multiple single RBCs before classifying. To classify multiple classes with deep learning, imbalance problems are common in medical imaging because normal samples are always higher than rare disease samples. This paper presents a new method to segment and classify RBCs from blood smear images, specifically to tackle cell overlapping and data imbalance problems. Focusing on overlapping cell separation, our segmentation process first estimates ellipses to represent RBCs. The method detects the concave points and then finds the ellipses using directed ellipse fitting. The accuracy from 20 blood smear images was 0.889. Classification requires balanced training datasets. However, some RBC types are rare. The imbalance ratio of this dataset was 34.538 for 12 RBC classes from 20,875 individual RBC samples. The use of machine learning for RBC classification with an imbalanced dataset is hence more challenging than many other applications. We analyzed techniques to deal with this problem. The best accuracy and F1-score were 0.921 and 0.8679, respectively, using EfficientNet-B1 with augmentation. Experimental results showed that the weight balancing technique with augmentation had the potential to deal with imbalance problems by improving the F1-score on minority classes, while data augmentation significantly improved the overall classification performance.
Deploying off-the-shelf segmentation networks on biomedical data has become common practice, yet if structures of interest in an image sequence are visible only temporarily, existing frame-by-frame methods fail. In this paper, we provide a solution to segmentation of imperfect data through time based on temporal propagation and uncertainty estimation. We integrate uncertainty estimation into Mask R-CNN network and propagate motion-corrected segmentation masks from frames with low uncertainty to those frames with high uncertainty to handle temporary loss of signal for segmentation. We demonstrate the value of this approach over frame-by-frame segmentation and regular temporal propagation on data from human embryonic kidney (HEK293T) cells transiently transfected with a fluorescent protein that moves in and out of the nucleus over time. The method presented here will empower microscopic experiments aimed at understanding molecular and cellular function.
Detecting and segmenting object instances is a common task in biomedical applications. Examples range from detecting lesions on functional magnetic resonance images, to the detection of tumours in histopathological images and extracting quantitative single-cell information from microscopy imagery, where cell segmentation is a major bottleneck. Attention-based transformers are state-of-the-art in a range of deep learning fields. They have recently been proposed for segmentation tasks where they are beginning to outperforming other methods. We present a novel attention-based cell detection transformer (Cell-DETR) for direct end-to-end instance segmentation. While the segmentation performance is on par with a state-of-the-art instance segmentation method, Cell-DETR is simpler and faster. We showcase the method's contribution in a the typical use case of segmenting yeast in microstructured environments, commonly employed in systems or synthetic biology. For the specific use case, the proposed method surpasses the state-of-the-art tools for semantic segmentation and additionally predicts the individual object instances. The fast and accurate instance segmentation performance increases the experimental information yield for a posteriori data processing and makes online monitoring of experiments and closed-loop optimal experimental design feasible.
Cell segmentation is a major bottleneck in extracting quantitative single-cell information from microscopy data. The challenge is exasperated in the setting of microstructured environments. While deep learning approaches have proven useful for general cell segmentation tasks, existing segmentation tools for the yeast-microstructure setting rely on traditional machine learning approaches. Here we present convolutional neural networks trained for multiclass segmenting of individual yeast cells and discerning these from cell-similar microstructures. We give an overview of the datasets recorded for training, validating and testing the networks, as well as a typical use-case. We showcase the method's contribution to segmenting yeast in microstructured environments with a typical synthetic biology application in mind. The models achieve robust segmentation results, outperforming the previous state-of-the-art in both accuracy and speed. The combination of fast and accurate segmentation is not only beneficial for a posteriori data processing, it also makes online monitoring of thousands of trapped cells or closed-loop optimal experimental design feasible from an image processing perspective.
Automatic analysis of spatio-temporal microscopy images is inevitable for state-of-the-art research in the life sciences. Recent developments in deep learning provide powerful tools for automatic analyses of such image data, but heavily depend on the amount and quality of provided training data to perform well. To this end, we developed a new method for realistic generation of synthetic 2D+t microscopy image data of fluorescently labeled cellular nuclei. The method combines spatiotemporal statistical shape models of different cell cycle stages with a conditional GAN to generate time series of cell populations and provides instance-level control of cell cycle stage and the fluorescence intensity of generated cells. We show the effect of the GAN conditioning and create a set of synthetic images that can be readily used for training and benchmarking of cell segmentation and tracking approaches.
Recent microscopy imaging techniques allow to precisely analyze cell morphology in 3D image data. To process the vast amount of image data generated by current digitized imaging techniques, automated approaches are demanded more than ever. Segmentation approaches used for morphological analyses, however, are often prone to produce unnaturally shaped predictions, which in conclusion could lead to inaccurate experimental outcomes. In order to minimize further manual interaction, shape priors help to constrain the predictions to the set of natural variations. In this paper, we show how spherical harmonics can be used as an alternative way to inherently constrain the predictions of neural networks for the segmentation of cells in 3D microscopy image data. Benefits and limitations of the spherical harmonic representation are analyzed and final results are compared to other state-of-the-art approaches on two different data sets.
Accurate labeling is essential for supervised deep learning methods. However, it is almost impossible to accurately and manually annotate thousands of images, which results in many labeling errors for most datasets. We proposes a local label point correction (LLPC) method to improve annotation quality for edge detection and image segmentation tasks. Our algorithm contains three steps: gradient-guided point correction, point interpolation and local point smoothing. We correct the labels of object contours by moving the annotated points to the pixel gradient peaks. This can improve the edge localization accuracy, but it also causes unsmooth contours due to the interference of image noise. Therefore, we design a point smoothing method based on local linear fitting to smooth the corrected edge. To verify the effectiveness of our LLPC, we construct a largest overlapping cervical cell edge detection dataset (CCEDD) with higher precision label corrected by our label correction method. Our LLPC only needs to set three parameters, but yields 30-40$\%$ average precision improvement on multiple networks. The qualitative and quantitative experimental results show that our LLPC can improve the quality of manual labels and the accuracy of overlapping cell edge detection. We hope that our study will give a strong boost to the development of the label correction for edge detection and image segmentation. We will release the dataset and code at https://github.com/nachifur/LLPC.
The impressive growth of data throughput in optical microscopy has triggered a widespread use of supervised learning (SL) models running on compressed image datasets for efficient automated analysis. However, since lossy image compression risks to produce unpredictable artifacts, quantifying the effect of data compression on SL applications is of pivotal importance to assess their reliability, especially for clinical use. We propose an experimental method to evaluate the tolerability of image compression distortions in 2D and 3D cell segmentation SL tasks: predictions on compressed data are compared to the raw predictive uncertainty, which is numerically estimated from the raw noise statistics measured through sensor calibration. We show that predictions on object- and image-specific segmentation parameters can be altered by up to 15% and more than 10 standard deviations after 16-to-8 bits downsampling or JPEG compression. In contrast, a recently developed lossless compression algorithm provides a prediction spread which is statistically equivalent to that stemming from raw noise, while providing a compression ratio of up to 10:1. By setting a lower bound to the SL predictive uncertainty, our technique can be generalized to validate a variety of data analysis pipelines in SL-assisted fields.
Recently, single-stage embedding based deep learning algorithms gain increasing attention in cell segmentation and tracking. Compared with the traditional "segment-then-associate" two-stage approach, a single-stage algorithm not only simultaneously achieves consistent instance cell segmentation and tracking but also gains superior performance when distinguishing ambiguous pixels on boundaries and overlaps. However, the deployment of an embedding based algorithm is restricted by slow inference speed (e.g., around 1-2 mins per frame). In this study, we propose a novel Faster Mean-shift algorithm, which tackles the computational bottleneck of embedding based cell segmentation and tracking. Different from previous GPU-accelerated fast mean-shift algorithms, a new online seed optimization policy (OSOP) is introduced to adaptively determine the minimal number of seeds, accelerate computation, and save GPU memory. With both embedding simulation and empirical validation via the four cohorts from the ISBI cell tracking challenge, the proposed Faster Mean-shift algorithm achieved 7-10 times speedup compared to the state-of-the-art embedding based cell instance segmentation and tracking algorithm. Our Faster Mean-shift algorithm also achieved the highest computational speed compared to other GPU benchmarks with optimized memory consumption. The Faster Mean-shift is a plug-and-play model, which can be employed on other pixel embedding based clustering inference for medical image analysis. (Plug-and-play model is publicly available: https://github.com/masqm/Faster-Mean-Shift)
Automatic cell segmentation in microscopy images works well with the support of deep neural networks trained with full supervision. Collecting and annotating images, though, is not a sustainable solution for every new microscopy database and cell type. Instead, we assume that we can access a plethora of annotated image data sets from different domains (sources) and a limited number of annotated image data sets from the domain of interest (target), where each domain denotes not only different image appearance but also a different type of cell segmentation problem. We pose this problem as meta-learning where the goal is to learn a generic and adaptable few-shot learning model from the available source domain data sets and cell segmentation tasks. The model can be afterwards fine-tuned on the few annotated images of the target domain that contains different image appearance and different cell type. In our meta-learning training, we propose the combination of three objective functions to segment the cells, move the segmentation results away from the classification boundary using cross-domain tasks, and learn an invariant representation between tasks of the source domains. Our experiments on five public databases show promising results from 1- to 10-shot meta-learning using standard segmentation neural network architectures.
Segmentation is a fundamental process in microscopic cell image analysis. With the advent of recent advances in deep learning, more accurate and high-throughput cell segmentation has become feasible. However, most existing deep learning-based cell segmentation algorithms require fully annotated ground-truth cell labels, which are time-consuming and labor-intensive to generate. In this paper, we introduce Scribble2Label, a novel weakly-supervised cell segmentation framework that exploits only a handful of scribble annotations without full segmentation labels. The core idea is to combine pseudo-labeling and label filtering to generate reliable labels from weak supervision. For this, we leverage the consistency of predictions by iteratively averaging the predictions to improve pseudo labels. We demonstrate the performance of Scribble2Label by comparing it to several state-of-the-art cell segmentation methods with various cell image modalities, including bright-field, fluorescence, and electron microscopy. We also show that our method performs robustly across different levels of scribble details, which confirms that only a few scribble annotations are required in real-use cases.
Deep Learning based methods have emerged as the indisputable leaders for virtually all image restoration tasks. Especially in the domain of microscopy images, various content-aware image restoration (CARE) approaches are now used to improve the interpretability of acquired data. Naturally, there are limitations to what can be restored in corrupted images, and like for all inverse problems, many potential solutions exist, and one of them must be chosen. Here, we propose DivNoising, a denoising approach based on fully convolutional variational autoencoders (VAEs), overcoming the problem of having to choose a single solution by predicting a whole distribution of denoised images. First we introduce a principled way of formulating the unsupervised denoising problem within the VAE framework by explicitly incorporating imaging noise models into the decoder. Our approach is fully unsupervised, only requiring noisy images and a suitable description of the imaging noise distribution. We show that such a noise model can either be measured, bootstrapped from noisy data, or co-learned during training. If desired, consensus predictions can be inferred from a set of DivNoising predictions, leading to competitive results with other unsupervised methods and, on occasion, even with the supervised state-of-the-art. DivNoising samples from the posterior enable a plethora of useful applications. We are (i) showing denoising results for 13 datasets, (ii) discussing how optical character recognition (OCR) applications can benefit from diverse predictions, and are (iii) demonstrating how instance cell segmentation improves when using diverse DivNoising predictions.
Semantic image segmentation is the process of labeling each pixel of an image with its corresponding class. An encoder-decoder based approach, like U-Net and its variants, is a popular strategy for solving medical image segmentation tasks. To improve the performance of U-Net on various segmentation tasks, we propose a novel architecture called DoubleU-Net, which is a combination of two U-Net architectures stacked on top of each other. The first U-Net uses a pre-trained VGG-19 as the encoder, which has already learned features from ImageNet and can be transferred to another task easily. To capture more semantic information efficiently, we added another U-Net at the bottom. We also adopt Atrous Spatial Pyramid Pooling (ASPP) to capture contextual information within the network. We have evaluated DoubleU-Net using four medical segmentation datasets, covering various imaging modalities such as colonoscopy, dermoscopy, and microscopy. Experiments on the MICCAI 2015 segmentation challenge, the CVC-ClinicDB, the 2018 Data Science Bowl challenge, and the Lesion boundary segmentation datasets demonstrate that the DoubleU-Net outperforms U-Net and the baseline models. Moreover, DoubleU-Net produces more accurate segmentation masks, especially in the case of the CVC-ClinicDB and MICCAI 2015 segmentation challenge datasets, which have challenging images such as smaller and flat polyps. These results show the improvement over the existing U-Net model. The encouraging results, produced on various medical image segmentation datasets, show that DoubleU-Net can be used as a strong baseline for both medical image segmentation and cross-dataset evaluation testing to measure the generalizability of Deep Learning (DL) models.
Detecting and segmenting individual cells from microscopy images is critical to various life science applications. Traditional cell segmentation tools are often ill-suited for applications in brightfield microscopy due to poor contrast and intensity heterogeneity, and only a small subset are applicable to segment cells in a cluster. In this regard, we introduce a novel supervised technique for cell segmentation in a multi-task learning paradigm. A combination of a multi-task loss, based on the region and cell boundary detection, is employed for an improved prediction efficiency of the network. The learning problem is posed in a novel min-max framework which enables adaptive estimation of the hyper-parameters in an automatic fashion. The region and cell boundary predictions are combined via morphological operations and active contour model to segment individual cells. The proposed methodology is particularly suited to segment touching cells from brightfield microscopy images without manual interventions. Quantitatively, we observe an overall Dice score of 0.93 on the validation set, which is an improvement of over 15.9% on a recent unsupervised method, and outperforms the popular supervised U-net algorithm by at least $5.8\%$ on average.