Spatio-temporal localization is vital for precise interactions across diverse domains, from biological research to autonomous navigation and interactive interfaces. Current video-based approaches, while proficient in tracking, lack the sophisticated reasoning capabilities of large language models, limiting their contextual understanding and generalization. We introduce VideoMolmo, a large multimodal model tailored for fine-grained spatio-temporal pointing conditioned on textual descriptions. Building upon the Molmo architecture, VideoMolmo incorporates a temporal module utilizing an attention mechanism to condition each frame on preceding frames, ensuring temporal consistency. Additionally, our novel temporal mask fusion pipeline employs SAM2 for bidirectional point propagation, significantly enhancing coherence across video sequences. This two-step decomposition, i.e., first using the LLM to generate precise pointing coordinates, then relying on a sequential mask-fusion module to produce coherent segmentation, not only simplifies the task for the language model but also enhances interpretability. Due to the lack of suitable datasets, we curate a comprehensive dataset comprising 72k video-caption pairs annotated with 100k object points. To evaluate the generalization of VideoMolmo, we introduce VPoS-Bench, a challenging out-of-distribution benchmark spanning five real-world scenarios: Cell Tracking, Egocentric Vision, Autonomous Driving, Video-GUI Interaction, and Robotics. We also evaluate our model on Referring Video Object Segmentation (Refer-VOS) and Reasoning VOS tasks. In comparison to existing models, VideoMolmo substantially improves spatio-temporal pointing accuracy and reasoning capability. Our code and models are publicly available at https://github.com/mbzuai-oryx/VideoMolmo.
Arabidopsis is a widely used model plant to gain basic knowledge on plant physiology and development. Live imaging is an important technique to visualize and quantify elemental processes in plant development. To uncover novel theories underlying plant growth and cell division, accurate cell tracking on live imaging is of utmost importance. The commonly used cell tracking software, TrackMate, adopts tracking-by-detection fashion, which applies Laplacian of Gaussian (LoG) for blob detection, and Linear Assignment Problem (LAP) tracker for tracking. However, they do not perform sufficiently when cells are densely arranged. To alleviate the problems mentioned above, we propose an accurate tracking method based on Genetic algorithm (GA) using knowledge of Arabidopsis root cellular patterns and spatial relationship among volumes. Our method can be described as a coarse-to-fine method, in which we first conducted relatively easy line-level tracking of cell nuclei, then performed complicated nuclear tracking based on known linear arrangement of cell files and their spatial relationship between nuclei. Our method has been evaluated on a long-time live imaging dataset of Arabidopsis root tips, and with minor manual rectification, it accurately tracks nuclei. To the best of our knowledge, this research represents the first successful attempt to address a long-standing problem in the field of time-lapse microscopy in the root meristem by proposing an accurate tracking method for Arabidopsis root nuclei.
Tracking individual cells in live-cell imaging provides fundamental insights, inevitable for studying causes and consequences of phenotypic heterogeneity, responses to changing environmental conditions or stressors. Microbial cell tracking, characterized by stochastic cell movements and frequent cell divisions, remains a challenging task when imaging frame rates must be limited to avoid counterfactual results. A promising way to overcome this limitation is uncertainty-aware tracking (UAT), which uses statistical models, calibrated to empirically observed cell behavior, to predict likely cell associations. We present PyUAT, an efficient and modular Python implementation of UAT for tracking microbial cells in time-lapse imaging. We demonstrate its performance on a large 2D+t data set and investigate the influence of modular biological models and imaging intervals on the tracking performance. The open-source PyUAT software is available at https://github.com/JuBiotech/PyUAT, including example notebooks for immediate use in Google Colab.
Cell tracking is a key computational task in live-cell microscopy, but fully automated analysis of high-throughput imaging requires reliable and, thus, uncertainty-aware data analysis tools, as the amount of data recorded within a single experiment exceeds what humans are able to overlook. We here propose and benchmark various methods to reason about and quantify uncertainty in linear assignment-based cell tracking algorithms. Our methods take inspiration from statistics and machine learning, leveraging two perspectives on the cell tracking problem explored throughout this work: Considering it as a Bayesian inference problem and as a classification problem. Our methods admit a framework-like character in that they equip any frame-to-frame tracking method with uncertainty quantification. We demonstrate this by applying it to various existing tracking algorithms including the recently presented Transformer-based trackers. We demonstrate empirically that our methods yield useful and well-calibrated tracking uncertainties.
Conventional multi-stage cell tracking approaches rely heavily on detection or segmentation in each frame as a prerequisite, requiring substantial resources for high-quality segmentation masks and increasing the overall prediction time. To address these limitations, we propose CAP, a novel end-to-end one-stage framework that reimagines cell tracking by treating Cell as Point. Unlike traditional methods, CAP eliminates the need for explicit detection or segmentation, instead jointly tracking cells for sequences in one stage by leveraging the inherent correlations among their trajectories. This simplification reduces both labeling requirements and pipeline complexity. However, directly processing the entire sequence in one stage poses challenges related to data imbalance in capturing cell division events and long sequence inference. To solve these challenges, CAP introduces two key innovations: (1) adaptive event-guided (AEG) sampling, which prioritizes cell division events to mitigate the occurrence imbalance of cell events, and (2) the rolling-as-window (RAW) inference strategy, which ensures continuous and stable tracking of newly emerging cells over extended sequences. By removing the dependency on segmentation-based preprocessing while addressing the challenges of imbalanced occurrence of cell events and long-sequence tracking, CAP demonstrates promising cell tracking performance and is 10 to 55 times more efficient than existing methods. The code and model checkpoints will be available soon.
Accurately tracking neuronal activity in behaving animals presents significant challenges due to complex motions and background noise. The lack of annotated datasets limits the evaluation and improvement of such tracking algorithms. To address this, we developed SINETRA, a versatile simulator that generates synthetic tracking data for particles on a deformable background, closely mimicking live animal recordings. This simulator produces annotated 2D and 3D videos that reflect the intricate movements seen in behaving animals like Hydra Vulgaris. We evaluated four state-of-the-art tracking algorithms highlighting the current limitations of these methods in challenging scenarios and paving the way for improved cell tracking techniques in dynamic biological systems.
Tracking the development of living cells in live-cell time-lapses reveals crucial insights into single-cell behavior and presents tremendous potential for biomedical and biotechnological applications. In microbial live-cell imaging (MLCI), a few to thousands of cells have to be detected and tracked within dozens of growing cell colonies. The challenge of tracking cells is heavily influenced by the experiment parameters, namely the imaging interval and maximal cell number. For now, tracking benchmarks are not widely available in MLCI and the effect of these parameters on the tracking performance are not yet known. Therefore, we present the largest publicly available and annotated dataset for MLCI, containing more than 1.4 million cell instances, 29k cell tracks, and 14k cell divisions. With this dataset at hand, we generalize existing tracking metrics to incorporate relevant imaging and experiment parameters into experiment-aware metrics. These metrics reveal that current cell tracking methods crucially depend on the choice of the experiment parameters, where their performance deteriorates at high imaging intervals and large cell colonies. Thus, our new benchmark quantifies the influence of experiment parameters on the tracking quality, and gives the opportunity to develop new data-driven methods that generalize across imaging and experiment parameters. The benchmark dataset is publicly available at https://zenodo.org/doi/10.5281/zenodo.7260136.
We introduce DynaCLR, a self-supervised framework for modeling cell dynamics via contrastive learning of representations of time-lapse datasets. Live cell imaging of cells and organelles is widely used to analyze cellular responses to perturbations. Human annotation of dynamic cell states captured by time-lapse perturbation datasets is laborious and prone to bias. DynaCLR integrates single-cell tracking with time-aware contrastive learning to map images of cells at neighboring time points to neighboring embeddings. Mapping the morphological dynamics of cells to a temporally regularized embedding space makes the annotation, classification, clustering, or interpretation of the cell states more quantitative and efficient. We illustrate the features and applications of DynaCLR with the following experiments: analyzing the kinetics of viral infection in human cells, detecting transient changes in cell morphology due to cell division, and mapping the dynamics of organelles due to viral infection. Models trained with DynaCLR consistently achieve $>95\%$ accuracy for infection state classification, enable the detection of transient cell states and reliably embed unseen experiments. DynaCLR provides a flexible framework for comparative analysis of cell state dynamics due to perturbations, such as infection, gene knockouts, and drugs. We provide PyTorch-based implementations of the model training and inference pipeline (https://github.com/mehta-lab/viscy) and a user interface (https://github.com/czbiohub-sf/napari-iohub) for the visualization and annotation of trajectories of cells in the real space and the embedding space.
The evaluation of cell tracking results steers the development of tracking methods, significantly impacting biomedical research. This is quantitatively achieved by means of evaluation metrics. Unfortunately, current metrics favor local correctness and weakly reward global coherence, impeding high-level biological analysis. To also foster global coherence, we propose the CHOTA metric (Cell-specific Higher Order Tracking Accuracy) which unifies the evaluation of all relevant aspects of cell tracking: cell detections and local associations, global coherence, and lineage tracking. We achieve this by introducing a new definition of the term 'trajectory' that includes the entire cell lineage and by including this into the well-established HOTA metric from general multiple object tracking. Furthermore, we provide a detailed survey of contemporary cell tracking metrics to compare our novel CHOTA metric and to show its advantages. All metrics are extensively evaluated on state-of-the-art real-data cell tracking results and synthetic results that simulate specific tracking errors. We show that CHOTA is sensitive to all tracking errors and gives a good indication of the biologically relevant capability of a method to reconstruct the full lineage of cells. It introduces a robust and comprehensive alternative to the currently used metrics in cell tracking. Python code is available at https://github.com/CellTrackingChallenge/py-ctcmetrics .
Live-cell microscopy allows to go beyond measuring average features of cellular populations to observe, quantify and explain biological heterogeneity. Deep Learning-based instance segmentation and cell tracking form the gold standard analysis tools to process the microscopy data collected, but tracking in particular suffers severely from low temporal resolution. In this work, we show that approximating cell cycle time distributions in microbial colonies of C. glutamicum is possible without performing tracking, even at low temporal resolution. To this end, we infer the parameters of a stochastic multi-stage birth process model using the Bayesian Synthetic Likelihood method at varying temporal resolutions by subsampling microscopy sequences, for which ground truth tracking is available. Our results indicate, that the proposed approach yields high quality approximations even at very low temporal resolution, where tracking fails to yield reasonable results.
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.
Generative models hold promise for revolutionizing medical education, robot-assisted surgery, and data augmentation for medical AI development. Diffusion models can now generate realistic images from text prompts, while recent advancements have demonstrated their ability to create diverse, high-quality videos. However, these models often struggle with generating accurate representations of medical procedures and detailed anatomical structures. This paper introduces Bora, the first spatio-temporal diffusion probabilistic model designed for text-guided biomedical video generation. Bora leverages Transformer architecture and is pre-trained on general-purpose video generation tasks. It is fine-tuned through model alignment and instruction tuning using a newly established medical video corpus, which includes paired text-video data from various biomedical fields. To the best of our knowledge, this is the first attempt to establish such a comprehensive annotated biomedical video dataset. Bora is capable of generating high-quality video data across four distinct biomedical domains, adhering to medical expert standards and demonstrating consistency and diversity. This generalist video generative model holds significant potential for enhancing medical consultation and decision-making, particularly in resource-limited settings. Additionally, Bora could pave the way for immersive medical training and procedure planning. Extensive experiments on distinct medical modalities such as endoscopy, ultrasound, MRI, and cell tracking validate the effectiveness of our model in understanding biomedical instructions and its superior performance across subjects compared to state-of-the-art generation models.
Automatic cell tracking in dense environments is plagued by inaccurate correspondences and misidentification of parent-offspring relationships. In this paper, we introduce a novel cell tracking algorithm named DenseTrack, which integrates deep learning with mathematical model-based strategies to effectively establish correspondences between consecutive frames and detect cell division events in crowded scenarios. We formulate the cell tracking problem as a deep learning-based temporal sequence classification task followed by solving a constrained one-to-one matching optimization problem exploiting the classifier's confidence scores. Additionally, we present an eigendecomposition-based cell division detection strategy that leverages knowledge of cellular geometry. The performance of the proposed approach has been evaluated by tracking densely packed cells in 3D time-lapse image sequences of bacterial biofilm development. The experimental results on simulated as well as experimental fluorescence image sequences suggest that the proposed tracking method achieves superior performance in terms of both qualitative and quantitative evaluation measures compared to recent state-of-the-art cell tracking approaches.
Cell tracking is a ubiquitous image analysis task in live-cell microscopy. Unlike multiple object tracking (MOT) for natural images, cell tracking typically involves hundreds of similar-looking objects that can divide in each frame, making it a particularly challenging problem. Current state-of-the-art approaches follow the tracking-by-detection paradigm, i.e. first all cells are detected per frame and successively linked in a second step to form biologically consistent cell tracks. Linking is commonly solved via discrete optimization methods, which require manual tuning of hyperparameters for each dataset and are therefore cumbersome to use in practice. Here we propose Trackastra, a general purpose cell tracking approach that uses a simple transformer architecture to directly learn pairwise associations of cells within a temporal window from annotated data. Importantly, unlike existing transformer-based MOT pipelines, our learning architecture also accounts for dividing objects such as cells and allows for accurate tracking even with simple greedy linking, thus making strides towards removing the requirement for a complex linking step. The proposed architecture operates on the full spatio-temporal context of detections within a time window by avoiding the computational burden of processing dense images. We show that our tracking approach performs on par with or better than highly tuned state-of-the-art cell tracking algorithms for various biological datasets, such as bacteria, cell cultures and fluorescent particles. We provide code at https://github.com/weigertlab/trackastra.
Cell tracking remains a pivotal yet challenging task in biomedical research. The full potential of deep learning for this purpose is often untapped due to the limited availability of comprehensive and varied training data sets. In this paper, we present SynCellFactory, a generative cell video augmentation. At the heart of SynCellFactory lies the ControlNet architecture, which has been fine-tuned to synthesize cell imagery with photorealistic accuracy in style and motion patterns. This technique enables the creation of synthetic yet realistic cell videos that mirror the complexity of authentic microscopy time-lapses. Our experiments demonstrate that SynCellFactory boosts the performance of well-established deep learning models for cell tracking, particularly when original training data is sparse.
Cell tracking and segmentation assist biologists in extracting insights from large-scale microscopy time-lapse data. Driven by local accuracy metrics, current tracking approaches often suffer from a lack of long-term consistency and the ability to reconstruct lineage trees correctly. To address this issue, we introduce an uncertainty estimation technique for motion estimation frameworks and extend the multi-hypothesis tracking framework. Our uncertainty estimation lifts motion representations into probabilistic spatial densities using problem-specific test-time augmentations. Moreover, we introduce a novel mitosis-aware assignment problem formulation that allows multi-hypothesis trackers to model cell splits and to resolve false associations and mitosis detections based on long-term conflicts. In our framework, explicit biological knowledge is modeled in assignment costs. We evaluate our approach on nine competitive datasets and demonstrate that we outperform the current state-of-the-art on biologically inspired metrics substantially, achieving improvements by a factor of approximately 6 and uncover new insights into the behavior of motion estimation uncertainty.
3D cell tracking in a living organism has a crucial role in live cell image analysis. Cell tracking in C. elegans has two difficulties. First, cell migration in a consecutive frame is large since they move their head during scanning. Second, cell detection is often inconsistent in consecutive frames due to touching cells and low-contrast images, and these inconsistent detections affect the tracking performance worse. In this paper, we propose a cell tracking method to address these issues, which has two main contributions. First, we introduce cell position heatmap-based non-rigid alignment with test-time fine-tuning, which can warp the detected points to near the positions at the next frame. Second, we propose a pairwise detection method, which uses the information of detection results at the previous frame for detecting cells at the current frame. The experimental results demonstrate the effectiveness of each module, and the proposed method achieved the best performance in comparison.
Determining cell identities in imaging sequences is an important yet challenging task. The conventional method for cell identification is via cell tracking, which is complex and can be time-consuming. In this study, we propose an innovative approach to cell identification during early $\textit{C. elegans}$ embryogenesis using machine learning. Cell identification during $\textit{C. elegans}$ embryogenesis would provide insights into neural development with implications for higher organisms including humans. We employed random forest, MLP, and LSTM models, and tested cell classification accuracy on 3D time-lapse confocal datasets spanning the first 4 hours of embryogenesis. By leveraging a small number of spatial-temporal features of individual cells, including cell trajectory and cell fate information, our models achieve an accuracy of over 91%, even with limited data. We also determine the most important feature contributions and can interpret these features in the context of biological knowledge. Our research demonstrates the success of predicting cell identities in time-lapse imaging sequences directly from simple spatio-temporal features.
Automatic detection and tracking of cells in microscopy images are major applications of computer vision technologies in both biomedical research and clinical practice. Though machine learning methods are increasingly common in these fields, classical algorithms still offer significant advantages for both tasks, including better explainability, faster computation, lower hardware requirements and more consistent performance. In this paper, we present a novel approach based on gravitational force fields that can compete with, and potentially outperform modern machine learning models when applied to fluorescence microscopy images. This method includes detection, segmentation, and tracking elements, with the results demonstrated on a Cell Tracking Challenge dataset.
Cell detection and tracking are paramount for bio-analysis. Recent approaches rely on the tracking-by-model evolution paradigm, which usually consists of training end-to-end deep learning models to detect and track the cells on the frames with promising results. However, such methods require extensive amounts of annotated data, which is time-consuming to obtain and often requires specialized annotators. This work proposes a new approach based on the classical tracking-by-detection paradigm that alleviates the requirement of annotated data. More precisely, it approximates the cell shapes as oriented ellipses and then uses generic-purpose oriented object detectors to identify the cells in each frame. We then rely on a global data association algorithm that explores temporal cell similarity using probability distance metrics, considering that the ellipses relate to two-dimensional Gaussian distributions. Our results show that our method can achieve detection and tracking results competitively with state-of-the-art techniques that require considerably more extensive data annotation. Our code is available at: https://github.com/LucasKirsten/Deep-Cell-Tracking-EBB.