Advanced image segmentation and processing tools present an opportunity to study cell processes and their dynamics. However, image analysis is often routine and time-consuming. Nowadays, alternative data-driven approaches using deep learning are potentially offering automatized, accurate, and fast image analysis. In this paper, we extend the applications of Cellpose, a state-of-the-art cell segmentation framework, with feature extraction capabilities to assess morphological characteristics. We also introduce a dataset of DAPI and FITC stained cells to which our new method is applied.
Time-lapse microscopy imaging is a crucial technique in biomedical studies for observing cellular behavior over time, providing essential data on cell numbers, sizes, shapes, and interactions. Manual analysis of hundreds or thousands of cells is impractical, necessitating the development of automated cell segmentation approaches. Traditional image processing methods have made significant progress in this area, but the advent of deep learning methods, particularly those using U-Net-based networks, has further enhanced performance in medical and microscopy image segmentation. However, challenges remain, particularly in accurately segmenting touching cells in images with low signal-to-noise ratios. Existing methods often struggle with effectively integrating features across different levels of abstraction. This can lead to model confusion, particularly when important contextual information is lost or the features are not adequately distinguished. The challenge lies in appropriately combining these features to preserve critical details while ensuring robust and accurate segmentation. To address these issues, we propose a novel framework called RA-SE-ASPP-Net, which incorporates Residual Blocks, Attention Mechanism, Squeeze-and-Excitation connection, and Atrous Spatial Pyramid Pooling to achieve precise and robust cell segmentation. We evaluate our proposed architecture using an induced pluripotent stem cell reprogramming dataset, a challenging dataset that has received limited attention in this field. Additionally, we compare our model with different ablation experiments to demonstrate its robustness. The proposed architecture outperforms the baseline models in all evaluated metrics, providing the most accurate semantic segmentation results. Finally, we applied the watershed method to the semantic segmentation results to obtain precise segmentations with specific information for each cell.
Histopathological imaging is vital for cancer research and clinical practice, with multiplexed Immunofluorescence (MxIF) and Hematoxylin and Eosin (H&E) providing complementary insights. However, aligning different stains at the cell level remains a challenge due to modality differences. In this paper, we present a novel framework for multimodal image alignment using cell segmentation outcomes. By treating cells as point sets, we apply Coherent Point Drift (CPD) for initial alignment and refine it with Graph Matching (GM). Evaluated on ovarian cancer tissue microarrays (TMAs), our method achieves high alignment accuracy, enabling integration of cell-level features across modalities and generating virtual H&E images from MxIF data for enhanced clinical interpretation.
Deep learning has revolutionized medical and biological imaging, particularly in segmentation tasks. However, segmenting biological cells remains challenging due to the high variability and complexity of cell shapes. Addressing this challenge requires high-quality datasets that accurately represent the diverse morphologies found in biological cells. Existing cell segmentation datasets are often limited by their focus on regular and uniform shapes. In this paper, we introduce a novel benchmark dataset of Ntera-2 (NT2) cells, a pluripotent carcinoma cell line, exhibiting diverse morphologies across multiple stages of differentiation, capturing the intricate and heterogeneous cellular structures that complicate segmentation tasks. To address these challenges, we propose an uncertainty-aware deep learning framework for complex cellular morphology segmentation (MorphoSeg) by incorporating sampling of virtual outliers from low-likelihood regions during training. Our comprehensive experimental evaluations against state-of-the-art baselines demonstrate that MorphoSeg significantly enhances segmentation accuracy, achieving up to a 7.74% increase in the Dice Similarity Coefficient (DSC) and a 28.36% reduction in the Hausdorff Distance. These findings highlight the effectiveness of our dataset and methodology in advancing cell segmentation capabilities, especially for complex and variable cell morphologies. The dataset and source code is publicly available at https://github.com/RanchoGoose/MorphoSeg.
Biomedical image segmentation is crucial for accurately diagnosing and analyzing various diseases. However, Convolutional Neural Networks (CNNs) and Transformers, the most commonly used architectures for this task, struggle to effectively capture long-range dependencies due to the inherent locality of CNNs and the computational complexity of Transformers. To address this limitation, we introduce TTT-Unet, a novel framework that integrates Test-Time Training (TTT) layers into the traditional U-Net architecture for biomedical image segmentation. TTT-Unet dynamically adjusts model parameters during the testing time, enhancing the model's ability to capture both local and long-range features. We evaluate TTT-Unet on multiple medical imaging datasets, including 3D abdominal organ segmentation in CT and MR images, instrument segmentation in endoscopy images, and cell segmentation in microscopy images. The results demonstrate that TTT-Unet consistently outperforms state-of-the-art CNN-based and Transformer-based segmentation models across all tasks. The code is available at https://github.com/rongzhou7/TTT-Unet.
Automated cell segmentation is crucial for various biological and medical applications, facilitating tasks like cell counting, morphology analysis, and drug discovery. However, manual segmentation is time-consuming and prone to subjectivity, necessitating robust automated methods. This paper presents open-source infrastructure, utilizing the UNet model, a deep-learning architecture noted for its effectiveness in image segmentation tasks. This implementation is integrated into the open-source DeepChem package, enhancing accessibility and usability for researchers and practitioners. The resulting tool offers a convenient and user-friendly interface, reducing the barrier to entry for cell segmentation while maintaining high accuracy. Additionally, we benchmark this model against various datasets, demonstrating its robustness and versatility across different imaging conditions and cell types.
Cell and nucleus segmentation are fundamental tasks for quantitative bioimage analysis. Despite progress in recent years, biologists and other domain experts still require novel algorithms to handle increasingly large and complex real-world datasets. These algorithms must not only achieve state-of-the-art accuracy, but also be optimized for efficiency, portability and user-friendliness. Here, we introduce InstanSeg: a novel embedding-based instance segmentation pipeline designed to identify cells and nuclei in microscopy images. Using six public cell segmentation datasets, we demonstrate that InstanSeg can significantly improve accuracy when compared to the most widely used alternative methods, while reducing the processing time by at least 60%. Furthermore, InstanSeg is designed to be fully serializable as TorchScript and supports GPU acceleration on a range of hardware. We provide an open-source implementation of InstanSeg in Python, in addition to a user-friendly, interactive QuPath extension for inference written in Java. Our code and pre-trained models are available at https://github.com/instanseg/instanseg .
In pathology, accurate and efficient analysis of Hematoxylin and Eosin (H\&E) slides is crucial for timely and effective cancer diagnosis. Although many deep learning solutions for nuclei instance segmentation and classification exist in the literature, they often entail high computational costs and resource requirements, thus limiting their practical usage in medical applications. To address this issue, we introduce a novel convolutional neural network, NuLite, a U-Net-like architecture designed explicitly on Fast-ViT, a state-of-the-art (SOTA) lightweight CNN. We obtained three versions of our model, NuLite-S, NuLite-M, and NuLite-H, trained on the PanNuke dataset. The experimental results prove that our models equal CellViT (SOTA) in terms of panoptic quality and detection. However, our lightest model, NuLite-S, is 40 times smaller in terms of parameters and about 8 times smaller in terms of GFlops, while our heaviest model is 17 times smaller in terms of parameters and about 7 times smaller in terms of GFlops. Moreover, our model is up to about 8 times faster than CellViT. Lastly, to prove the effectiveness of our solution, we provide a robust comparison of external datasets, namely CoNseP, MoNuSeg, and GlySAC. Our model is publicly available at https://github.com/CosmoIknosLab/NuLite
Lensless fiber endomicroscope is an emerging tool for in-vivo microscopic imaging, where quantitative phase imaging (QPI) can be utilized as a label-free method to enhance image contrast. However, existing single-shot phase reconstruction methods through lensless fiber endomicroscope typically perform well on simple images but struggle with complex microscopic structures. Here, we propose a speckle-conditioned diffusion model (SpecDiffusion), which reconstructs phase images directly from speckles captured at the detection side of a multi-core fiber (MCF). Unlike conventional neural networks, SpecDiffusion employs iterative phase denoising steps for speckle-driven phase reconstruction. The iteration scheme allows SpecDiffusion to break down the phase reconstruction process into multiple steps, gradually building up to the final phase image. This attribute alleviates the computation challenge at each step and enables the reconstruction of rich details in complex microscopic images. To validate its efficacy, we build an optical system to capture speckles from MCF and construct a dataset consisting of 100,000 paired images. SpecDiffusion provides high-fidelity phase reconstruction results and shows powerful generalization capacity for unseen objects, such as test charts and biological tissues, reducing the average mean absolute error of the reconstructed tissue images by 7 times. Furthermore, the reconstructed tissue images using SpecDiffusion shows higher accuracy in zero-shot cell segmentation tasks compared to the conventional method, demonstrating the potential for further cell morphology analysis through the learning-based lensless fiber endomicroscope. SpecDiffusion offers a precise and generalized method to phase reconstruction through scattering media, including MCFs, opening new perspective in lensless fiber endomicroscopic imaging.
Immunofluorescent (IF) imaging is crucial for visualizing biomarker expressions, cell morphology and assessing the effects of drug treatments on sub-cellular components. IF imaging needs extra staining process and often requiring cell fixation, therefore it may also introduce artefects and alter endogenouous cell morphology. Some IF stains are expensive or not readily available hence hindering experiments. Recent diffusion models, which synthesise high-fidelity IF images from easy-to-acquire brightfield (BF) images, offer a promising solution but are hindered by training instability and slow inference times due to the noise diffusion process. This paper presents a novel method for the conditional synthesis of IF images directly from BF images along with cell segmentation masks. Our approach employs a Residual Diffusion process that enhances stability and significantly reduces inference time. We performed a critical evaluation against other image-to-image synthesis models, including UNets, GANs, and advanced diffusion models. Our model demonstrates significant improvements in image quality (p<0.05 in MSE, PSNR, and SSIM), inference speed (26 times faster than competing diffusion models), and accurate segmentation results for both nuclei and cell bodies (0.77 and 0.63 mean IOU for nuclei and cell true positives, respectively). This paper is a substantial advancement in the field, providing robust and efficient tools for cell image analysis.
The precise segmentation and tracking of cells in microscopy image sequences play a pivotal role in biomedical research, facilitating the study of tissue, organ, and organism development. However, manual segmentation and tracking of cells is time-consuming and often require professional experiences. Besides, segmenting cells in the images with a low signal-to-noise ratio remains difficult. While deep learning (DL) has become a common method for cell segmentation, few DL- based methods address concurrent cell segmentation and tracking. In this paper, we propose a novel DL approach featuring graph-based tracking for cell segmentation and tracking in microscopy images. We combine Deeplabv3+ for semantic segmentation and ResNet50 for enhanced feature extraction, enabling comprehensive cell detection and instance segmentation. Post-processing, involving non-maxima suppression and outlier detection, refines predictions and produces final segmentation. The tracking method is based on the relative position of graph nodes to track segmented cells, encompassing cell division and apoptosis. We conduct our experiments on the induced pluripotent stem (iPS) cell datasets, and the results show that the segmentation and tracking performance of our method yields superior performance compared to the benchmark models. More specifically, our approach achieved DET values of 0.955 and 0.913, TRA values of 0.951 and 0.906, and SEG values of 0.690 and 0.665 on two iPS dataset videos, respectively. Additionally, the performance assessment encompasses four real microscopy datasets from the Cell Tracking Challenge (CTC). Our method greatly reduces the cost of manual labeling and labor-intensive costs.
The segmentation of cell nuclei in tissue images stained with the blood dye hematoxylin and eosin (H$\&$E) is essential for various clinical applications and analyses. Due to the complex characteristics of cellular morphology, a large receptive field is considered crucial for generating high-quality segmentation. However, previous methods face challenges in achieving a balance between the receptive field and computational burden. To address this issue, we propose LKCell, a high-accuracy and efficient cell segmentation method. Its core insight lies in unleashing the potential of large convolution kernels to achieve computationally efficient large receptive fields. Specifically, (1) We transfer pre-trained large convolution kernel models to the medical domain for the first time, demonstrating their effectiveness in cell segmentation. (2) We analyze the redundancy of previous methods and design a new segmentation decoder based on large convolution kernels. It achieves higher performance while significantly reducing the number of parameters. We evaluate our method on the most challenging benchmark and achieve state-of-the-art results (0.5080 mPQ) in cell nuclei instance segmentation with only 21.6% FLOPs compared with the previous leading method. Our source code and models are available at https://github.com/hustvl/LKCell.
Accurately segmenting and individualizing cells in SEM images is a highly promising technique for elucidating tissue architecture in oncology. While current AI-based methods are effective, errors persist, necessitating time-consuming manual corrections, particularly in areas where the quality of cell contours in the image is poor and requires gap filling. This study presents a novel AI-driven approach for refining cell boundary delineation to improve instance-based cell segmentation in SEM images, also reducing the necessity for residual manual correction. A CNN COp-Net is introduced to address gaps in cell contours, effectively filling in regions with deficient or absent information. The network takes as input cell contour probability maps with potentially inadequate or missing information and outputs corrected cell contour delineations. The lack of training data was addressed by generating low integrity probability maps using a tailored PDE. We showcase the efficacy of our approach in augmenting cell boundary precision using both private SEM images from PDX hepatoblastoma tissues and publicly accessible images datasets. The proposed cell contour closing operator exhibits a notable improvement in tested datasets, achieving respectively close to 50% (private data) and 10% (public data) increase in the accurately-delineated cell proportion compared to state-of-the-art methods. Additionally, the need for manual corrections was significantly reduced, therefore facilitating the overall digitalization process. Our results demonstrate a notable enhancement in the accuracy of cell instance segmentation, particularly in highly challenging regions where image quality compromises the integrity of cell boundaries, necessitating gap filling. Therefore, our work should ultimately facilitate the study of tumour tissue bioarchitecture in onconanotomy field.
Early detection and accurate diagnosis can predict the risk of malignant disease transformation, thereby increasing the probability of effective treatment. Identifying mild syndrome with small pathological regions serves as an ominous warning and is fundamental in the early diagnosis of diseases. While deep learning algorithms, particularly convolutional neural networks (CNNs), have shown promise in segmenting medical objects, analyzing small areas in medical images remains challenging. This difficulty arises due to information losses and compression defects from convolution and pooling operations in CNNs, which become more pronounced as the network deepens, especially for small medical objects. To address these challenges, we propose a novel scale-variant attention-based network (SvANet) for accurately segmenting small-scale objects in medical images. The SvANet consists of scale-variant attention, cross-scale guidance, Monte Carlo attention, and vision transformer, which incorporates cross-scale features and alleviates compression artifacts for enhancing the discrimination of small medical objects. Quantitative experimental results demonstrate the superior performance of SvANet, achieving 96.12%, 96.11%, 89.79%, 84.15%, 80.25%, 73.05%, and 72.58% in mean Dice coefficient for segmenting kidney tumors, skin lesions, hepatic tumors, polyps, surgical excision cells, retinal vasculatures, and sperms, which occupy less than 1% of the image areas in KiTS23, ISIC 2018, ATLAS, PolypGen, TissueNet, FIVES, and SpermHealth datasets, respectively.
Accurate segmentation of medical images is crucial for diagnostic purposes, including cell segmentation, tumor identification, and organ localization. Traditional convolutional neural network (CNN)-based approaches struggled to achieve precise segmentation results due to their limited receptive fields, particularly in cases involving multi-organ segmentation with varying shapes and sizes. The transformer-based approaches address this limitation by leveraging the global receptive field, but they often face challenges in capturing local information required for pixel-precise segmentation. In this work, we introduce DwinFormer, a hierarchical encoder-decoder architecture for medical image segmentation comprising a directional window (Dwin) attention and global self-attention (GSA) for feature encoding. The focus of our design is the introduction of Dwin block within DwinFormer that effectively captures local and global information along the horizontal, vertical, and depthwise directions of the input feature map by separately performing attention in each of these directional volumes. To this end, our Dwin block introduces a nested Dwin attention (NDA) that progressively increases the receptive field in horizontal, vertical, and depthwise directions and a convolutional Dwin attention (CDA) that captures local contextual information for the attention computation. While the proposed Dwin block captures local and global dependencies at the first two high-resolution stages of DwinFormer, the GSA block encodes global dependencies at the last two lower-resolution stages. Experiments over the challenging 3D Synapse Multi-organ dataset and Cell HMS dataset demonstrate the benefits of our DwinFormer over the state-of-the-art approaches. Our source code will be publicly available at \url{https://github.com/Daniyanaj/DWINFORMER}.
Deep Learning (DL) models have been successfully applied to many applications including biomedical cell segmentation and classification in histological images. These models require large amounts of annotated data which might not always be available, especially in the medical field where annotations are scarce and expensive. To overcome this limitation, we propose a novel pipeline for generating synthetic datasets for cell segmentation. Given only a handful of annotated images, our method generates a large dataset of images which can be used to effectively train DL instance segmentation models. Our solution is designed to generate cells of realistic shapes and placement by allowing experts to incorporate domain knowledge during the generation of the dataset.
Automated cell segmentation has become increasingly crucial for disease diagnosis and drug discovery, as manual delineation is excessively laborious and subjective. To address this issue with limited manual annotation, researchers have developed semi/unsupervised segmentation approaches. Among these approaches, the Deep Gaussian mixture model plays a vital role due to its capacity to facilitate complex data distributions. However, these models assume that the data follows symmetric normal distributions, which is inapplicable for data that is asymmetrically distributed. These models also obstacles weak generalization capacity and are sensitive to outliers. To address these issues, this paper presents a novel asymmetric mixture model for unsupervised cell segmentation. This asymmetric mixture model is built by aggregating certain multivariate Gaussian mixture models with log-likelihood and self-supervised-based optimization functions. The proposed asymmetric mixture model outperforms (nearly 2-30% gain in dice coefficient, p<0.05) the existing state-of-the-art unsupervised models on cell segmentation including the segment anything.
Purpose: Quantitative phase imaging (QPI) is a label-free technique that provides high-contrast images of tissues and cells without the use of chemicals or dyes. Accurate semantic segmentation of cells in QPI is essential for various biomedical applications. While DM-based segmentation has demonstrated promising results, the requirement for multiple sampling steps reduces efficiency. This study aims to enhance DM-based segmentation by introducing prior-guided content information into the starting noise, thereby minimizing inefficiencies associated with multiple sampling. Approach: A prior-guided mechanism is introduced into DM-based segmentation, replacing randomly sampled starting noise with noise informed by content information. This mechanism utilizes another trained DM and DDIM inversion to incorporate content information from the to-be-segmented images into the starting noise. An evaluation method is also proposed to assess the quality of the starting noise, considering both content and distribution information. Results: Extensive experiments on various QPI datasets for cell segmentation showed that the proposed method achieved superior performance in DM-based segmentation with only a single sampling. Ablation studies and visual analysis further highlighted the significance of content priors in DM-based segmentation. Conclusion: The proposed method effectively leverages prior content information to improve DM-based segmentation, providing accurate results while reducing the need for multiple samplings. The findings emphasize the importance of integrating content priors into DM-based segmentation methods for optimal performance.
Cell image segmentation is usually implemented using fully supervised deep learning methods, which heavily rely on extensive annotated training data. Yet, due to the complexity of cell morphology and the requirement for specialized knowledge, pixel-level annotation of cell images has become a highly labor-intensive task. To address the above problems, we propose an active learning framework for cell segmentation using bounding box annotations, which greatly reduces the data annotation cost of cell segmentation algorithms. First, we generate a box-supervised learning method (denoted as YOLO-SAM) by combining the YOLOv8 detector with the Segment Anything Model (SAM), which effectively reduces the complexity of data annotation. Furthermore, it is integrated into an active learning framework that employs the MC DropBlock method to train the segmentation model with fewer box-annotated samples. Extensive experiments demonstrate that our model saves more than ninety percent of data annotation time compared to mask-supervised deep learning methods.
Cell segmentation is essential in biomedical research for analyzing cellular morphology and behavior. Deep learning methods, particularly convolutional neural networks (CNNs), have revolutionized cell segmentation by extracting intricate features from images. However, the robustness of these methods under microscope optical aberrations remains a critical challenge. This study evaluates cell image segmentation models under optical aberrations from fluorescence and bright field microscopy. By simulating different types of aberrations, including astigmatism, coma, spherical aberration, trefoil, and mixed aberrations, we conduct a thorough evaluation of various cell instance segmentation models using the DynamicNuclearNet (DNN) and LIVECell datasets, representing fluorescence and bright field microscopy cell datasets, respectively. We train and test several segmentation models, including the Otsu threshold method and Mask R-CNN with different network heads (FPN, C3) and backbones (ResNet, VGG, Swin Transformer), under aberrated conditions. Additionally, we provide usage recommendations for the Cellpose 2.0 Toolbox on complex cell degradation images. The results indicate that the combination of FPN and SwinS demonstrates superior robustness in handling simple cell images affected by minor aberrations. In contrast, Cellpose 2.0 proves effective for complex cell images under similar conditions. Furthermore, we innovatively propose the Point Spread Function Image Label Classification Model (PLCM). This model can quickly and accurately identify aberration types and amplitudes from PSF images, assisting researchers without optical training. Through PLCM, researchers can better apply our proposed cell segmentation guidelines.