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.
Cell segmentation is a critical step for quantitative single-cell analysis in microscopy images. Existing cell segmentation methods are often tailored to specific modalities or require manual interventions to specify hyper-parameters in different experimental settings. Here, we present a multi-modality cell segmentation benchmark, comprising over 1500 labeled images derived from more than 50 diverse biological experiments. The top participants developed a Transformer-based deep-learning algorithm that not only exceeds existing methods but can also be applied to diverse microscopy images across imaging platforms and tissue types without manual parameter adjustments. This benchmark and the improved algorithm offer promising avenues for more accurate and versatile cell analysis in microscopy imaging.
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/ .
Cell nuclei segmentation is challenging due to shape variation and closely clustered or partially overlapping objects. Most previous methods are not globally optimal, limited to elliptical models, or are computationally expensive. In this work, we introduce a globally optimal approach based on deformable shape models and global energy minimization for cell nuclei segmentation and cluster splitting. We propose an implicit parameterization of deformable shape models and show that it leads to a convex energy. Convex energy minimization yields the global solution independently of the initialization, is fast, and robust. To jointly perform cell nuclei segmentation and cluster splitting, we developed a novel iterative global energy minimization method, which leverages the inherent property of superadditivity of the convex energy. This property exploits the lower bound of the energy of the union of the models and improves the computational efficiency. Our method provably determines a solution close to global optimality. In addition, we derive a closed-form solution of the proposed global minimization based on the superadditivity property for non-clustered cell nuclei. We evaluated our method using fluorescence microscopy images of five different cell types comprising various challenges, and performed a quantitative comparison with previous methods. Our method achieved state-of-the-art or improved performance.
Although deep learning (DL) shows powerful potential in cell segmentation tasks, it suffers from poor generalization as DL-based methods originally simplified cell segmentation in detecting cell membrane boundary, lacking prominent cellular structures to position overall differentiating. Moreover, the scarcity of annotated cell images limits the performance of DL models. Segmentation limitations of a single category of cell make massive practice difficult, much less, with varied modalities. In this paper, we introduce a novel semi-supervised cell segmentation method called Multi-Microscopic-view Cell semi-supervised Segmentation (MMCS), which can train cell segmentation models utilizing less labeled multi-posture cell images with different microscopy well. Technically, MMCS consists of Nucleus-assisted global recognition, Self-adaptive diameter filter, and Temporal-ensembling models. Nucleus-assisted global recognition adds additional cell nucleus channel to improve the global distinguishing performance of fuzzy cell membrane boundaries even when cells aggregate. Besides, self-adapted cell diameter filter can help separate multi-resolution cells with different morphology properly. It further leverages the temporal-ensembling models to improve the semi-supervised training process, achieving effective training with less labeled data. Additionally, optimizing the weight of unlabeled loss contributed to total loss also improve the model performance. Evaluated on the Tuning Set of NeurIPS 2022 Cell Segmentation Challenge (NeurIPS CellSeg), MMCS achieves an F1-score of 0.8239 and the running time for all cases is within the time tolerance.
Mapping spatial distributions of transcriptomic cell types is essential to understanding the brain, with its exceptional cellular heterogeneity and the functional significance of its spatial organization. Spatial transcriptomics techniques are hoped to accomplish these measurements, but each method uses different experimental and computational protocols, with different trade-offs and optimizations. In 2017, the SpaceTx Consortium was formed to compare these methods and determine their suitability for large-scale spatial transcriptomic atlases. SpaceTx work included progress in tissue processing, taxonomy development, gene selection, image processing and data standardization, cell segmentation, cell type assignments, and visualization. Although the landscape of experimental methods has changed dramatically since the beginning of SpaceTx, the need for quantitative and detailed benchmarking of spatial transcriptomics methods in the brain is still unmet. Here, we summarize the work of SpaceTx and highlight outstanding challenges as spatial transcriptomics grows into a mature field. We also discuss how our progress provides a roadmap for benchmarking spatial transcriptomics methods in the future. Data and analyses from this consortium, along with code and methods are publicly available at https://spacetx.github.io/.
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
Accurate segmentation of live cell images has broad applications in clinical and research contexts. Deep learning methods have been able to perform cell segmentations with high accuracy; however developing machine learning models to do this requires access to high fidelity images of live cells. This is often not available due to resource constraints like limited accessibility to high performance microscopes or due to the nature of the studied organisms. Segmentation on low resolution images of live cells is a difficult task. This paper proposes a method to perform live cell segmentation with low resolution images by performing super-resolution as a pre-processing step in the segmentation pipeline.
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.
Melanoma is one of the most aggressive forms of skin cancer, causing a large proportion of skin cancer deaths. However, melanoma diagnoses by pathologists shows low interrater reliability. As melanoma is a cancer of the melanocyte, there is a clear need to develop a melanocytic cell segmentation tool that is agnostic to pathologist variability and automates pixel-level annotation. Gigapixel-level pathologist labeling, however, is impractical. Herein, we propose a means to train deep neural networks for melanocytic cell segmentation from hematoxylin and eosin (H&E) stained sections and paired immunohistochemistry (IHC) of adjacent tissue sections, achieving a mean IOU of 0.64 despite imperfect ground-truth labels.
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.