Our work targets automated analysis to quantify the growth dynamics of a population of bacilliform bacteria. We propose an innovative approach to frame-sequence tracking of deformable-cell motion by the automated minimization of a new, specific cost functional. This minimization is implemented by dedicated Boltzmann machines (stochastic recurrent neural networks). Automated detection of cell divisions is handled similarly by successive minimizations of two cost functions, alternating the identification of children pairs and parent identification. We validate the proposed automatic cell tracking algorithm using (i) recordings of simulated cell colonies that closely mimic the growth dynamics of E. coli in microfluidic traps and (ii) real data. On a batch of 1100 simulated image frames, cell registration accuracies per frame ranged from 94.5% to 100%, with a high average. Our initial tests using experimental image sequences (i.e., real data) of E. coli colonies also yield convincing results, with a registration accuracy ranging from 90% to 100%.
Cell segmentation and tracking in microscopy images are of great significance to new discoveries in biology and medicine. In this study, we propose a novel approach to combine cell segmentation and cell tracking into a unified end-to-end deep learning based framework, where cell detection and segmentation are performed with a current instance segmentation pipeline and cell tracking is implemented by integrating Siamese Network with the pipeline. Besides, tracking performance is improved by incorporating spatial information into the network and fusing spatial and visual prediction. Our approach was evaluated on the DeepCell benchmark dataset. Despite being simple and efficient, our method outperforms state-of-the-art algorithms in terms of both cell segmentation and cell tracking accuracies.
Multi-object tracking (MOT) in computer vision and cell tracking in biomedical image analysis are two similar research fields, whose common aim is to achieve instance level object detection/segmentation and associate such objects across different video frames. However, one major difference between these two tasks is that cell tracking also aim to detect mitosis (cell division), which is typically not considered in MOT tasks. Therefore, the acyclic oriented graphs matching (AOGM) has been used as de facto standard evaluation metrics for cell tracking, rather than directly using the evaluation metrics in computer vision, such as multiple object tracking accuracy (MOTA), ID Switches (IDS), ID F1 Score (IDF1) etc. However, based on our experiments, we realized that AOGM did not always function as expected for mitosis events. In this paper, we exhibit the limitations of evaluating mitosis with AOGM using both simulated and real cell tracking data.
Oblique back-illumination capillaroscopy has recently been introduced as a method for high-quality, non-invasive blood cell imaging in human capillaries. To make this technique practical for clinical blood cell counting, solutions for automatic processing of acquired videos are needed. Here, we take the first step towards this goal, by introducing a deep learning multi-cell tracking model, named CycleTrack, which achieves accurate blood cell counting from capillaroscopic videos. CycleTrack combines two simple online tracking models, SORT and CenterTrack, and is tailored to features of capillary blood cell flow. Blood cells are tracked by displacement vectors in two opposing temporal directions (forward- and backward-tracking) between consecutive frames. This approach yields accurate tracking despite rapidly moving and deforming blood cells. The proposed model outperforms other baseline trackers, achieving 65.57% Multiple Object Tracking Accuracy and 73.95% ID F1 score on test videos. Compared to manual blood cell counting, CycleTrack achieves 96.58 $\pm$ 2.43% cell counting accuracy among 8 test videos with 1000 frames each compared to 93.45% and 77.02% accuracy for independent CenterTrack and SORT almost without additional time expense. It takes 800s to track and count approximately 8000 blood cells from 9,600 frames captured in a typical one-minute video. Moreover, the blood cell velocity measured by CycleTrack demonstrates a consistent, pulsatile pattern within the physiological range of heart rate. Lastly, we discuss future improvements for the CycleTrack framework, which would enable clinical translation of the oblique back-illumination microscope towards a real-time and non-invasive point-of-care blood cell counting and analyzing technology.
Analyzing the dynamical properties of mobile objects requires to extract trajectories from recordings, which is often done by tracking movies. We compiled a database of two-dimensional movies for very different biological and physical systems spanning a wide range of length scales and developed a general-purpose, optimized, open-source, cross-platform, easy to install and use, self-updating software called FastTrack. It can handle a changing number of deformable objects in a region of interest, and is particularly suitable for animal and cell tracking in two-dimensions. Furthermore, we introduce the probability of incursions as a new measure of a movie's trackability that doesn't require the knowledge of ground truth trajectories, since it is resilient to small amounts of errors and can be computed on the basis of an ad hoc tracking. We also leveraged the versatility and speed of FastTrack to implement an iterative algorithm determining a set of nearly-optimized tracking parameters -- yet further reducing the amount of human intervention -- and demonstrate that FastTrack can be used to explore the space of tracking parameters to optimize the number of swaps for a batch of similar movies. A benchmark shows that FastTrack is orders of magnitude faster than state-of-the-art tracking algorithms, with a comparable tracking accuracy. The source code is available under the GNU GPLv3 at https://github.com/FastTrackOrg/FastTrack and pre-compiled binaries for Windows, Mac and Linux are available at http://www.fasttrack.sh.
We propose a novel weakly supervised method to improve the boundary of the 3D segmented nuclei utilizing an over-segmented image. This is motivated by the observation that current state-of-the-art deep learning methods do not result in accurate boundaries when the training data is weakly annotated. Towards this, a 3D U-Net is trained to get the centroid of the nuclei and integrated with a simple linear iterative clustering (SLIC) supervoxel algorithm that provides better adherence to cluster boundaries. To track these segmented nuclei, our algorithm utilizes the relative nuclei location depicting the processes of nuclei division and apoptosis. The proposed algorithmic pipeline achieves better segmentation performance compared to the state-of-the-art method in Cell Tracking Challenge (CTC) 2019 and comparable performance to state-of-the-art methods in IEEE ISBI CTC2020 while utilizing very few pixel-wise annotated data. Detailed experimental results are provided, and the source code is available on GitHub.
Motivation: Innovative microfluidic systems carry the promise to greatly facilitate spatio-temporal analysis of single cells under well-defined environmental conditions, allowing novel insights into population heterogeneity and opening new opportunities for fundamental and applied biotechnology. Microfluidics experiments, however, are accompanied by vast amounts of data, such as time series of microscopic images, for which manual evaluation is infeasible due to the sheer number of samples. While classical image processing technologies do not lead to satisfactory results in this domain, modern deep learning technologies such as convolutional networks can be sufficiently versatile for diverse tasks, including automatic cell tracking and counting as well as the extraction of critical parameters, such as growth rate. However, for successful training, current supervised deep learning requires label information, such as the number or positions of cells for each image in a series; obtaining these annotations is very costly in this setting. Results: We propose a novel Machine Learning architecture together with a specialized training procedure, which allows us to infuse a deep neural network with human-powered abstraction on the level of data, leading to a high-performing regression model that requires only a very small amount of labeled data. Specifically, we train a generative model simultaneously on natural and synthetic data, so that it learns a shared representation, from which a target variable, such as the cell count, can be reliably estimated.
Multiple Object Tracking (MOT) detects the trajectories of multiple objects given an input video. It has become more and more important for various research and industry areas, such as cell tracking for biomedical research and human tracking in video surveillance. Most existing algorithms depend on the uniqueness of the object's appearance, and the dominating bipartite matching scheme ignores the speed smoothness. Although several methods have incorporated the velocity smoothness for tracking, they either fail to pursue global smooth velocity or are often trapped in local optimums. We focus on the general MOT problem regardless of the appearance and propose an appearance-free tripartite matching to avoid the irregular velocity problem of the bipartite matching. The tripartite matching is formulated as maximizing the likelihood of the state vectors constituted of the position and velocity of objects, which results in a chain-dependent structure. We resort to the dynamic programming algorithm to find such a maximum likelihood estimate. To overcome the high computational cost induced by the vast search space of dynamic programming when many objects are to be tracked, we decompose the space by the number of disappearing objects and propose a reduced-space approach by truncating the decomposition. Extensive simulations have shown the superiority and efficiency of our proposed method, and the comparisons with top methods on Cell Tracking Challenge also demonstrate our competence. We also applied our method to track the motion of natural killer cells around tumor cells in a cancer study.\footnote{The source code is available on \url{https://github.com/szcf-weiya/TriMatchMOT}
We propose a weakly-supervised cell tracking method that can train a convolutional neural network (CNN) by using only the annotation of "cell detection" (i.e., the coordinates of cell positions) without association information, in which cell positions can be easily obtained by nuclear staining. First, we train co-detection CNN that detects cells in successive frames by using weak-labels. Our key assumption is that co-detection CNN implicitly learns association in addition to detection. To obtain the association, we propose a backward-and-forward propagation method that analyzes the correspondence of cell positions in the outputs of co-detection CNN. Experiments demonstrated that the proposed method can associate cells by analyzing co-detection CNN. Even though the method uses only weak supervision, the performance of our method was almost the same as the state-of-the-art supervised method. Code is publicly available in https://github.com/naivete5656/WSCTBFP
Recently, single-stage embedding based deep learning algorithms gain increasing attention in cell segmentation and tracking. Compared with the traditional "segment-then-associate" two-stage approach, a single-stage algorithm not only simultaneously achieves consistent instance cell segmentation and tracking but also gains superior performance when distinguishing ambiguous pixels on boundaries and overlaps. However, the deployment of an embedding based algorithm is restricted by slow inference speed (e.g., around 1-2 mins per frame). In this study, we propose a novel Faster Mean-shift algorithm, which tackles the computational bottleneck of embedding based cell segmentation and tracking. Different from previous GPU-accelerated fast mean-shift algorithms, a new online seed optimization policy (OSOP) is introduced to adaptively determine the minimal number of seeds, accelerate computation, and save GPU memory. With both embedding simulation and empirical validation via the four cohorts from the ISBI cell tracking challenge, the proposed Faster Mean-shift algorithm achieved 7-10 times speedup compared to the state-of-the-art embedding based cell instance segmentation and tracking algorithm. Our Faster Mean-shift algorithm also achieved the highest computational speed compared to other GPU benchmarks with optimized memory consumption. The Faster Mean-shift is a plug-and-play model, which can be employed on other pixel embedding based clustering inference for medical image analysis. (Plug-and-play model is publicly available: https://github.com/masqm/Faster-Mean-Shift)
State-of-the-art multi-object tracking~(MOT) methods follow the tracking-by-detection paradigm, where object trajectories are obtained by associating per-frame outputs of object detectors. In crowded scenes, however, detectors often fail to obtain accurate detections due to heavy occlusions and high crowd density. In this paper, we propose a new MOT paradigm, tracking-by-counting, tailored for crowded scenes. Using crowd density maps, we jointly model detection, counting, and tracking of multiple targets as a network flow program, which simultaneously finds the global optimal detections and trajectories of multiple targets over the whole video. This is in contrast to prior MOT methods that either ignore the crowd density and thus are prone to errors in crowded scenes, or rely on a suboptimal two-step process using heuristic density-aware point-tracks for matching targets.Our approach yields promising results on public benchmarks of various domains including people tracking, cell tracking, and fish tracking.
We propose a weakly-supervised cell tracking method that can train a convolutional neural network (CNN) by using only the annotation of "cell detection" (i.e., the coordinates of cell positions) without association information, in which cell positions can be easily obtained by nuclear staining. First, we train a co-detection CNN that detects cells in successive frames by using weak-labels. Our key assumption is that the co-detection CNN implicitly learns association in addition to detection. To obtain the association information, we propose a backward-and-forward propagation method that analyzes the correspondence of cell positions in the detection maps output of the co-detection CNN. Experiments demonstrated that the proposed method can match positions by analyzing the co-detection CNN. Even though the method uses only weak supervision, the performance of our method was almost the same as the state-of-the-art supervised method.
We propose a fast approximate solver for the combinatorial problem known as tracking-by-assignment, which we apply to cell tracking. The latter plays a key role in discovery in many life sciences, especially in cell and developmental biology. So far, in the most general setting this problem was addressed by off-the-shelf solvers like Gurobi, whose run time and memory requirements rapidly grow with the size of the input. In contrast, for our method this growth is nearly linear. Our contribution consists of a new (1) decomposable compact representation of the problem; (2) dual block-coordinate ascent method for optimizing the decomposition-based dual; and (3) primal heuristics that reconstructs a feasible integer solution based on the dual information. Compared to solving the problem with Gurobi, we observe an up to~60~times speed-up, while reducing the memory footprint significantly. We demonstrate the efficacy of our method on real-world tracking problems.
We propose a cell segmentation method for analyzing images of densely clustered cells. The method combines the strengths of marker-controlled watershed transformation and a convolutional neural network (CNN). We demonstrate the method universality and high performance on three Cell Tracking Challenge (CTC) datasets of clustered cells captured by different acquisition techniques. For all tested datasets, our method reached the top performance in both cell detection and segmentation. Based on a series of experiments, we observed: (1) Predicting both watershed marker function and segmentation function significantly improves the accuracy of the segmentation. (2) Both functions can be learned independently. (3) Training data augmentation by scaling and rigid geometric transformations is superior to augmentation that involves elastic transformations. Our method is simple to use, and it generalizes well for various data with state-of-the-art performance.
The accurate segmentation and tracking of cells in microscopy image sequences is an important task in biomedical research, e.g., for studying the development of tissues, organs or entire organisms. However, the segmentation of touching cells in images with a low signal-to-noise-ratio is still a challenging problem. In this paper, we present a method for the segmentation of touching cells in microscopy images. By using a novel representation of cell borders, inspired by distance maps, our method is capable to utilize not only touching cells but also close cells in the training process. Furthermore, this representation is notably robust to annotation errors and shows promising results for the segmentation of microscopy images containing in the training data underrepresented or not included cell types. For the prediction of the proposed neighbor distances, an adapted U-Net convolutional neural network (CNN) with two decoder paths is used. In addition, we adapt a graph-based cell tracking algorithm to evaluate our proposed method on the task of cell tracking. The adapted tracking algorithm includes a movement estimation in the cost function to re-link tracks with missing segmentation masks over a short sequence of frames. Our combined tracking by detection method has proven its potential in the IEEE ISBI 2020 Cell Tracking Challenge (http://celltrackingchallenge.net/) where we achieved as team KIT-Sch-GE multiple top three rankings including two top performances using a single segmentation model for the diverse data sets.
Conventional cell tracking methods detect multiple cells in each frame (detection) and then associate the detection results in successive time-frames (association). Most cell tracking methods perform the association task independently from the detection task. However, there is no guarantee of preserving coherence between these tasks, and lack of coherence may adversely affect tracking performance. In this paper, we propose the Motion and Position Map (MPM) that jointly represents both detection and association for not only migration but also cell division. It guarantees coherence such that if a cell is detected, the corresponding motion flow can always be obtained. It is a simple but powerful method for multi-object tracking in dense environments. We compared the proposed method with current tracking methods under various conditions in real biological images and found that it outperformed the state-of-the-art (+5.2\% improvement compared to the second-best).
The ability to extrapolate gene expression dynamics in living single cells requires robust cell segmentation, and one of the challenges is the amorphous or irregularly shaped cell boundaries. To address this issue, we modified the U-Net architecture to segment cells in fluorescence widefield microscopy images and quantitatively evaluated its performance. We also proposed a novel loss function approach that emphasizes the segmentation accuracy on cell boundaries and encourages shape feature preservation. With a 97% sensitivity, 93% specificity, 91% Jaccard similarity, and 95% Dice coefficient, our proposed method called Residual Attention U-Net with edge-enhancement surpassed the state-of-the-art U-Net in segmentation performance as evaluated by the traditional metrics. More remarkably, the same proposed candidate also performed the best in terms of the preservation of valuable shape features, namely area, eccentricity, major axis length, solidity and orientation. These improvements on shape feature preservation can serve as useful assets for downstream cell tracking and quantification of changes in cell statistics or features over time.
Phase contrast microscopy (PCM) has been widely used in biomedicine research, which allows users to observe objectives without staining or killing them. One important related research is to employ PCM to monitor live cells. How to segment cell populations in obtained PCM images gains more and more attention as its a critical step for downstream applications, such as cell tracking, cell classification and others. Many papers have been published to deal with this problem from different perspectives. In this paper we aim to present a comprehensive review on the development of PCM cell population segmentation.
U-Net and its variants have been demonstrated to work sufficiently well in biological cell tracking and segmentation. However, these methods still suffer in the presence of complex processes such as collision of cells, mitosis and apoptosis. In this paper, we augment U-Net with Siamese matching-based tracking and propose to track individual nuclei over time. By modelling the behavioural pattern of the cells, we achieve improved segmentation and tracking performances through a re-segmentation procedure. Our preliminary investigations on the Fluo-N2DH-SIM+ and Fluo-N2DH-GOWT1 datasets demonstrate that absolute improvements of up to 3.8 % and 3.4% can be obtained in segmentation and tracking accuracy, respectively.
Tracking cell particles in 3D microscopy videos is a challenging task but is of great significance for modeling the motion of cells. Proper characterization of the cell's shape, evolution, and their movement over time is crucial to understanding and modeling the mechanobiology of cell migration in many diseases. One in particular, toxoplasmosis is the disease caused by the parasite Toxoplasma gondii. Roughly, one-third of the world's population tests positive for T. gondii. Its virulence is linked to its lytic cycle, predicated on its motility and ability to enter and exit nucleated cells; therefore, studies elucidating its motility patterns are critical to the eventual development of therapeutic strategies. Here, we present a computational framework for fast and scalable detection, tracking, and identification of T. gondii motion phenotypes in 3D videos, in a completely unsupervised fashion. Our pipeline consists of several different modules including preprocessing, sparsification, cell detection, cell tracking, trajectories extraction, parametrization of the trajectories; and finally, a clustering step. Additionally, we identified the computational bottlenecks, and developed a lightweight and highly scalable pipeline through a combination of task distribution and parallelism. Our results prove both the accuracy and performance of our method.