The segmentation and tracking of living cells play a vital role within the biomedical domain, particularly in cancer research, drug development, and developmental biology. These are usually tedious and time-consuming tasks that are traditionally done by biomedical experts. Recently, to automatize these processes, deep learning based segmentation and tracking methods have been proposed. These methods require large-scale datasets and their full potential is constrained by the scarcity of annotated data in the biomedical imaging domain. To address this limitation, we propose Biomedical Video Diffusion Model (BVDM), capable of generating realistic-looking synthetic microscopy videos. Trained only on a single real video, BVDM can generate videos of arbitrary length with pixel-level annotations that can be used for training data-hungry models. It is composed of a denoising diffusion probabilistic model (DDPM) generating high-fidelity synthetic cell microscopy images and a flow prediction model (FPM) predicting the non-rigid transformation between consecutive video frames. During inference, initially, the DDPM imposes realistic cell textures on synthetic cell masks which are generated based on real data statistics. The flow prediction model predicts the flow field between consecutive masks and applies that to the DDPM output from the previous time frame to create the next one while keeping temporal consistency. BVDM outperforms state-of-the-art synthetic live cell microscopy video generation models. Furthermore, we demonstrate that a sufficiently large synthetic dataset enhances the performance of cell segmentation and tracking models compared to using a limited amount of available real data.
Background: Cell segmentation in bright-field histological slides is a crucial topic in medical image analysis. Having access to accurate segmentation allows researchers to examine the relationship between cellular morphology and clinical observations. Unfortunately, most segmentation methods known today are limited to nuclei and cannot segmentate the cytoplasm. Material & Methods: We present a new network architecture Cyto R-CNN that is able to accurately segment whole cells (with both the nucleus and the cytoplasm) in bright-field images. We also present a new dataset CytoNuke, consisting of multiple thousand manual annotations of head and neck squamous cell carcinoma cells. Utilizing this dataset, we compared the performance of Cyto R-CNN to other popular cell segmentation algorithms, including QuPath's built-in algorithm, StarDist and Cellpose. To evaluate segmentation performance, we calculated AP50, AP75 and measured 17 morphological and staining-related features for all detected cells. We compared these measurements to the gold standard of manual segmentation using the Kolmogorov-Smirnov test. Results: Cyto R-CNN achieved an AP50 of 58.65% and an AP75 of 11.56% in whole-cell segmentation, outperforming all other methods (QuPath $19.46/0.91\%$; StarDist $45.33/2.32\%$; Cellpose $31.85/5.61\%$). Cell features derived from Cyto R-CNN showed the best agreement to the gold standard ($\bar{D} = 0.15$) outperforming QuPath ($\bar{D} = 0.22$), StarDist ($\bar{D} = 0.25$) and Cellpose ($\bar{D} = 0.23$). Conclusion: Our newly proposed Cyto R-CNN architecture outperforms current algorithms in whole-cell segmentation while providing more reliable cell measurements than any other model. This could improve digital pathology workflows, potentially leading to improved diagnosis. Moreover, our published dataset can be used to develop further models in the future.
Cell segmentation in histopathological images plays a crucial role in understanding, diagnosing, and treating many diseases. However, data annotation for this is expensive since there can be a large number of cells per image, and expert pathologists are needed for labelling images. Instead, our paper focuses on using weak supervision -- annotation from related tasks -- to induce a segmenter. Recent foundation models, such as Segment Anything (SAM), can use prompts to leverage additional supervision during inference. SAM has performed remarkably well in natural image segmentation tasks; however, its applicability to cell segmentation has not been explored. In response, we investigate guiding the prompting procedure in SAM for weakly supervised cell segmentation when only bounding box supervision is available. We develop two workflows: (1) an object detector's output as a test-time prompt to SAM (D-SAM), and (2) SAM as pseudo mask generator over training data to train a standalone segmentation model (SAM-S). On finding that both workflows have some complementary strengths, we develop an integer programming-based approach to reconcile the two sets of segmentation masks, achieving yet higher performance. We experiment on three publicly available cell segmentation datasets namely, ConSep, MoNuSeg, and TNBC, and find that all SAM-based solutions hugely outperform existing weakly supervised image segmentation models, obtaining 9-15 pt Dice gains.
Cell segmentation for multi-modal microscopy images remains a challenge due to the complex textures, patterns, and cell shapes in these images. To tackle the problem, we first develop an automatic cell classification pipeline to label the microscopy images based on their low-level image characteristics, and then train a classification model based on the category labels. Afterward, we train a separate segmentation model for each category using the images in the corresponding category. Besides, we further deploy two types of segmentation models to segment cells with roundish and irregular shapes respectively. Moreover, an efficient and powerful backbone model is utilized to enhance the efficiency of our segmentation model. Evaluated on the Tuning Set of NeurIPS 2022 Cell Segmentation Challenge, our method achieves an F1-score of 0.8795 and the running time for all cases is within the time tolerance.
Image segmentation plays an essential role in nuclei image analysis. Recently, the segment anything model has made a significant breakthrough in such tasks. However, the current model exists two major issues for cell segmentation: (1) the image encoder of the segment anything model involves a large number of parameters. Retraining or even fine-tuning the model still requires expensive computational resources. (2) in point prompt mode, points are sampled from the center of the ground truth and more than one set of points is expected to achieve reliable performance, which is not efficient for practical applications. In this paper, a single-point prompt network is proposed for nuclei image segmentation, called SPPNet. We replace the original image encoder with a lightweight vision transformer. Also, an effective convolutional block is added in parallel to extract the low-level semantic information from the image and compensate for the performance degradation due to the small image encoder. We propose a new point-sampling method based on the Gaussian kernel. The proposed model is evaluated on the MoNuSeg-2018 dataset. The result demonstrated that SPPNet outperforms existing U-shape architectures and shows faster convergence in training. Compared to the segment anything model, SPPNet shows roughly 20 times faster inference, with 1/70 parameters and computational cost. Particularly, only one set of points is required in both the training and inference phases, which is more reasonable for clinical applications. The code for our work and more technical details can be found at https://github.com/xq141839/SPPNet.
The success of supervised deep learning models on cell recognition tasks relies on detailed annotations. Many previous works have managed to reduce the dependency on labels. However, considering the large number of cells contained in a patch, costly and inefficient labeling is still inevitable. To this end, we explored label-free methods for cell recognition. Prior self-activation maps (PSM) are proposed to generate pseudo masks as training targets. To be specific, an activation network is trained with self-supervised learning. The gradient information in the shallow layers of the network is aggregated to generate prior self-activation maps. Afterward, a semantic clustering module is then introduced as a pipeline to transform PSMs to pixel-level semantic pseudo masks for downstream tasks. We evaluated our method on two histological datasets: MoNuSeg (cell segmentation) and BCData (multi-class cell detection). Compared with other fully-supervised and weakly-supervised methods, our method can achieve competitive performance without any manual annotations. Our simple but effective framework can also achieve multi-class cell detection which can not be done by existing unsupervised methods. The results show the potential of PSMs that might inspire other research to deal with the hunger for labels in medical area.
Eosinophilic esophagitis (EoE) is a chronic and relapsing disease characterized by esophageal inflammation. Symptoms of EoE include difficulty swallowing, food impaction, and chest pain which significantly impact the quality of life, resulting in nutritional impairments, social limitations, and psychological distress. The diagnosis of EoE is typically performed with a threshold (15 to 20) of eosinophils (Eos) per high-power field (HPF). Since the current counting process of Eos is a resource-intensive process for human pathologists, automatic methods are desired. Circle representation has been shown as a more precise, yet less complicated, representation for automatic instance cell segmentation such as CircleSnake approach. However, the CircleSnake was designed as a single-label model, which is not able to deal with multi-label scenarios. In this paper, we propose the multi-label CircleSnake model for instance segmentation on Eos. It extends the original CircleSnake model from a single-label design to a multi-label model, allowing segmentation of multiple object types. Experimental results illustrate the CircleSnake model's superiority over the traditional Mask R-CNN model and DeepSnake model in terms of average precision (AP) in identifying and segmenting eosinophils, thereby enabling enhanced characterization of EoE. This automated approach holds promise for streamlining the assessment process and improving diagnostic accuracy in EoE analysis. The source code has been made publicly available at https://github.com/yilinliu610730/EoE.
In this work, we describe a method for large-scale 3D cell-tracking through a segmentation selection approach. The proposed method is effective at tracking cells across large microscopy datasets on two fronts: (i) It can solve problems containing millions of segmentation instances in terabyte-scale 3D+t datasets; (ii) It achieves competitive results with or without deep learning, which requires 3D annotated data, that is scarce in the fluorescence microscopy field. The proposed method computes cell tracks and segments using a hierarchy of segmentation hypotheses and selects disjoint segments by maximizing the overlap between adjacent frames. We show that this method achieves state-of-the-art results in 3D images from the cell tracking challenge and has a faster integer linear programming formulation. Moreover, our framework is flexible and supports segmentations from off-the-shelf cell segmentation models and can combine them into an ensemble that improves tracking. The code is available https://github.com/royerlab/ultrack.
Annotating medical imaging datasets is costly, so fine-tuning (or transfer learning) is the most effective method for digital pathology vision applications such as disease classification and semantic segmentation. However, due to texture bias in models trained on real-world images, transfer learning for histopathology applications might result in underperforming models, which necessitates the need for using unlabeled histopathology data and self-supervised methods to discover domain-specific characteristics. Here, we tested the premise that histopathology-specific pretrained models provide better initializations for pathology vision tasks, i.e., gland and cell segmentation. In this study, we compare the performance of gland and cell segmentation tasks with histopathology domain-specific and non-domain-specific (real-world images) pretrained weights. Moreover, we investigate the dataset size at which domain-specific pretraining produces significant gains in performance. In addition, we investigated whether domain-specific initialization improves the effectiveness of out-of-distribution testing on distinct datasets but the same task. The results indicate that performance gain using domain-specific pretrained weights depends on both the task and the size of the training dataset. In instances with limited dataset sizes, a significant improvement in gland segmentation performance was also observed, whereas models trained on cell segmentation datasets exhibit no improvement.
Nuclei detection and segmentation in hematoxylin and eosin-stained (H&E) tissue images are important clinical tasks and crucial for a wide range of applications. However, it is a challenging task due to nuclei variances in staining and size, overlapping boundaries, and nuclei clustering. While convolutional neural networks have been extensively used for this task, we explore the potential of Transformer-based networks in this domain. Therefore, we introduce a new method for automated instance segmentation of cell nuclei in digitized tissue samples using a deep learning architecture based on Vision Transformer called CellViT. CellViT is trained and evaluated on the PanNuke dataset, which is one of the most challenging nuclei instance segmentation datasets, consisting of nearly 200,000 annotated Nuclei into 5 clinically important classes in 19 tissue types. We demonstrate the superiority of large-scale in-domain and out-of-domain pre-trained Vision Transformers by leveraging the recently published Segment Anything Model and a ViT-encoder pre-trained on 104 million histological image patches - achieving state-of-the-art nuclei detection and instance segmentation performance on the PanNuke dataset with a mean panoptic quality of 0.50 and an F1-detection score of 0.83. The code is publicly available at https://github.com/TIO-IKIM/CellViT
Current state-of-the-art supervised deep learning-based segmentation approaches have demonstrated superior performance in medical image segmentation tasks. However, such supervised approaches require fully annotated pixel-level ground-truth labels, which are labor-intensive and time-consuming to acquire. Recently, Scribble2Label (S2L) demonstrated that using only a handful of scribbles with self-supervised learning can generate accurate segmentation results without full annotation. However, owing to the relatively small size of scribbles, the model is prone to overfit and the results may be biased to the selection of scribbles. In this work, we address this issue by employing a novel multiscale contrastive regularization term for S2L. The main idea is to extract features from intermediate layers of the neural network for contrastive loss so that structures at various scales can be effectively separated. To verify the efficacy of our method, we conducted ablation studies on well-known datasets, such as Data Science Bowl 2018 and MoNuSeg. The results show that the proposed multiscale contrastive loss is effective in improving the performance of S2L, which is comparable to that of the supervised learning segmentation method.
Multi-class cell segmentation in high-resolution Giga-pixel whole slide images (WSI) is critical for various clinical applications. Training such an AI model typically requires labor-intensive pixel-wise manual annotation from experienced domain experts (e.g., pathologists). Moreover, such annotation is error-prone when differentiating fine-grained cell types (e.g., podocyte and mesangial cells) via the naked human eye. In this study, we assess the feasibility of democratizing pathological AI deployment by only using lay annotators (annotators without medical domain knowledge). The contribution of this paper is threefold: (1) We proposed a molecular-empowered learning scheme for multi-class cell segmentation using partial labels from lay annotators; (2) The proposed method integrated Giga-pixel level molecular-morphology cross-modality registration, molecular-informed annotation, and molecular-oriented segmentation model, so as to achieve significantly superior performance via 3 lay annotators as compared with 2 experienced pathologists; (3) A deep corrective learning (learning with imperfect label) method is proposed to further improve the segmentation performance using partially annotated noisy data. From the experimental results, our learning method achieved F1 = 0.8496 using molecular-informed annotations from lay annotators, which is better than conventional morphology-based annotations (F1 = 0.7015) from experienced pathologists. Our method democratizes the development of a pathological segmentation deep model to the lay annotator level, which consequently scales up the learning process similar to a non-medical computer vision task. The official implementation and cell annotations are publicly available at https://github.com/hrlblab/MolecularEL.
This paper presents advancements in automated early-stage prediction of the success of reprogramming human induced pluripotent stem cells (iPSCs) as a potential source for regenerative cell therapies.The minuscule success rate of iPSC-reprogramming of around $ 0.01% $ to $ 0.1% $ makes it labor-intensive, time-consuming, and exorbitantly expensive to generate a stable iPSC line. Since that requires culturing of millions of cells and intense biological scrutiny of multiple clones to identify a single optimal clone. The ability to reliably predict which cells are likely to establish as an optimal iPSC line at an early stage of pluripotency would therefore be ground-breaking in rendering this a practical and cost-effective approach to personalized medicine. Temporal information about changes in cellular appearance over time is crucial for predicting its future growth outcomes. In order to generate this data, we first performed continuous time-lapse imaging of iPSCs in culture using an ultra-high resolution microscope. We then annotated the locations and identities of cells in late-stage images where reliable manual identification is possible. Next, we propagated these labels backwards in time using a semi-automated tracking system to obtain labels for early stages of growth. Finally, we used this data to train deep neural networks to perform automatic cell segmentation and classification. Our code and data are available at https://github.com/abhineet123/ipsc_prediction.
Quantifying the phagocytosis of dynamic, unstained cells is essential for evaluating neurodegenerative diseases. However, measuring rapid cell interactions and distinguishing cells from background make this task very challenging when processing time-lapse phase-contrast video microscopy. In this study, we introduce an end-to-end, scalable, and versatile real-time framework for quantifying and analyzing phagocytic activity. Our proposed pipeline is able to process large data-sets and includes a data quality verification module to counteract perturbations such as microscope movements and frame blurring. We also propose an explainable cell segmentation module to improve the interpretability of DL methods compared to black-box algorithms. This includes two interpretable DL capabilities: visual explanation and model simplification. We demonstrate that interpretability in DL is not the opposite of high performance, by additionally providing essential DL algorithm optimization insights and solutions. Besides, incorporating interpretable modules results in an efficient architecture design and optimized execution time. We apply our pipeline to analyze microglial cell phagocytosis in FTD and obtain statistically reliable results showing that FTD mutant cells are larger and more aggressive than control cells. The method has been tested and validated on public benchmarks by generating state-of-the art performances. To stimulate translational approaches and future studies, we release an open-source end-to-end pipeline and a unique microglial cells phagocytosis dataset for immune system characterization in neurodegenerative diseases research. This pipeline and the associated dataset will consistently crystallize future advances in this field, promoting the development of interpretable algorithms dedicated to the domain of neurodegenerative diseases' characterization. github.com/ounissimehdi/PhagoStat
Despite their superior performance, deep-learning methods often suffer from the disadvantage of needing large-scale well-annotated training data. In response, recent literature has seen a proliferation of efforts aimed at reducing the annotation burden. This paper focuses on a weakly-supervised training setting for single-cell segmentation models, where the only available training label is the rough locations of individual cells. The specific problem is of practical interest due to the widely available nuclei counter-stain data in biomedical literature, from which the cell locations can be derived programmatically. Of more general interest is a proposed self-learning method called collaborative knowledge sharing, which is related to but distinct from the more well-known consistency learning methods. This strategy achieves self-learning by sharing knowledge between a principal model and a very light-weight collaborator model. Importantly, the two models are entirely different in their architectures, capacities, and model outputs: In our case, the principal model approaches the segmentation problem from an object-detection perspective, whereas the collaborator model a sematic segmentation perspective. We assessed the effectiveness of this strategy by conducting experiments on LIVECell, a large single-cell segmentation dataset of bright-field images, and on A431 dataset, a fluorescence image dataset in which the location labels are generated automatically from nuclei counter-stain data. Implementing code is available at https://github.com/jiyuuchc/lacss
Extracting single-cell information from microscopy data requires accurate instance-wise segmentations. Obtaining pixel-wise segmentations from microscopy imagery remains a challenging task, especially with the added complexity of microstructured environments. This paper presents a novel dataset for segmenting yeast cells in microstructures. We offer pixel-wise instance segmentation labels for both cells and trap microstructures. In total, we release 493 densely annotated microscopy images. To facilitate a unified comparison between novel segmentation algorithms, we propose a standardized evaluation strategy for our dataset. The aim of the dataset and evaluation strategy is to facilitate the development of new cell segmentation approaches. The dataset is publicly available at https://christophreich1996.github.io/yeast_in_microstructures_dataset/ .
Deep learning provides us with powerful methods to perform nucleus or cell segmentation with unprecedented quality. However, these methods usually require large training sets of manually annotated images, which are tedious and expensive to generate. In this paper we propose to use In Silico Labeling (ISL) as a pretraining scheme for segmentation tasks. The strategy is to acquire label-free microscopy images (such as bright-field or phase contrast) along fluorescently labeled images (such as DAPI or CellMask). We then train a model to predict the fluorescently labeled images from the label-free microscopy images. By comparing segmentation performance across several training set sizes, we show that such a scheme can dramatically reduce the number of required annotations.
Cell segmentation is a fundamental task for computational biology analysis. Identifying the cell instances is often the first step in various downstream biomedical studies. However, many cell segmentation algorithms, including the recently emerging deep learning-based methods, still show limited generality under the multi-modality environment. Weakly Supervised Cell Segmentation in Multi-modality High-Resolution Microscopy Images was hosted at NeurIPS 2022 to tackle this problem. We propose MEDIAR, a holistic pipeline for cell instance segmentation under multi-modality in this challenge. MEDIAR harmonizes data-centric and model-centric approaches as the learning and inference strategies, achieving a 0.9067 F1-score at the validation phase while satisfying the time budget. To facilitate subsequent research, we provide the source code and trained model as open-source: https://github.com/Lee-Gihun/MEDIAR
In microscopy image cell segmentation, it is common to train a deep neural network on source data, containing different types of microscopy images, and then fine-tune it using a support set comprising a few randomly selected and annotated training target images. In this paper, we argue that the random selection of unlabelled training target images to be annotated and included in the support set may not enable an effective fine-tuning process, so we propose a new approach to optimise this image selection process. Our approach involves a new scoring function to find informative unlabelled target images. In particular, we propose to measure the consistency in the model predictions on target images against specific data augmentations. However, we observe that the model trained with source datasets does not reliably evaluate consistency on target images. To alleviate this problem, we propose novel self-supervised pretext tasks to compute the scores of unlabelled target images. Finally, the top few images with the least consistency scores are added to the support set for oracle (i.e., expert) annotation and later used to fine-tune the model to the target images. In our evaluations that involve the segmentation of five different types of cell images, we demonstrate promising results on several target test sets compared to the random selection approach as well as other selection approaches, such as Shannon's entropy and Monte-Carlo dropout.
Single-cell technologies are revolutionizing the entire field of biology. The large volumes of data generated by single-cell technologies are high-dimensional, sparse, heterogeneous, and have complicated dependency structures, making analyses using conventional machine learning approaches challenging and impractical. In tackling these challenges, deep learning often demonstrates superior performance compared to traditional machine learning methods. In this work, we give a comprehensive survey on deep learning in single-cell analysis. We first introduce background on single-cell technologies and their development, as well as fundamental concepts of deep learning including the most popular deep architectures. We present an overview of the single-cell analytic pipeline pursued in research applications while noting divergences due to data sources or specific applications. We then review seven popular tasks spanning through different stages of the single-cell analysis pipeline, including multimodal integration, imputation, clustering, spatial domain identification, cell-type deconvolution, cell segmentation, and cell-type annotation. Under each task, we describe the most recent developments in classical and deep learning methods and discuss their advantages and disadvantages. Deep learning tools and benchmark datasets are also summarized for each task. Finally, we discuss the future directions and the most recent challenges. This survey will serve as a reference for biologists and computer scientists, encouraging collaborations.