scholarly journals Accurate cell tracking and lineage construction in live-cell imaging experiments with deep learning

2019 ◽  
Author(s):  
Erick Moen ◽  
Enrico Borba ◽  
Geneva Miller ◽  
Morgan Schwartz ◽  
Dylan Bannon ◽  
...  

AbstractLive-cell imaging experiments have opened an exciting window into the behavior of living systems. While these experiments can produce rich data, the computational analysis of these datasets is challenging. Single-cell analysis requires that cells be accurately identified in each image and subsequently tracked over time. Increasingly, deep learning is being used to interpret microscopy image with single cell resolution. In this work, we apply deep learning to the problem of tracking single cells in live-cell imaging data. Using crowdsourcing and a human-in-the-loop approach to data annotation, we constructed a dataset of over 11,000 trajectories of cell nuclei that includes lineage information. Using this dataset, we successfully trained a deep learning model to perform cell tracking within a linear programming framework. Benchmarking tests demonstrate that our method achieves state-of-the-art performance on the task of cell tracking with respect to multiple accuracy metrics. Further, we show that our deep learning-based method generalizes to perform cell tracking for both fluorescent and brightfield images of the cell cytoplasm, despite having never been trained on those data types. This enables analysis of live-cell imaging data collected across imaging modalities. A persistent cloud deployment of our cell tracker is available at http://www.deepcell.org.

2017 ◽  
Vol 4 (8) ◽  
pp. 170811 ◽  
Author(s):  
Sayak Mukherjee ◽  
David Stewart ◽  
William Stewart ◽  
Lewis L. Lanier ◽  
Jayajit Das

Single-cell responses are shaped by the geometry of signalling kinetic trajectories carved in a multidimensional space spanned by signalling protein abundances. It is, however, challenging to assay a large number (more than 3) of signalling species in live-cell imaging, which makes it difficult to probe single-cell signalling kinetic trajectories in large dimensions. Flow and mass cytometry techniques can measure a large number (4 to more than 40) of signalling species but are unable to track single cells. Thus, cytometry experiments provide detailed time-stamped snapshots of single-cell signalling kinetics. Is it possible to use the time-stamped cytometry data to reconstruct single-cell signalling trajectories? Borrowing concepts of conserved and slow variables from non-equilibrium statistical physics we develop an approach to reconstruct signalling trajectories using snapshot data by creating new variables that remain invariant or vary slowly during the signalling kinetics. We apply this approach to reconstruct trajectories using snapshot data obtained from in silico simulations, live-cell imaging measurements, and, synthetic flow cytometry datasets. The application of invariants and slow variables to reconstruct trajectories provides a radically different way to track objects using snapshot data. The approach is likely to have implications for solving matching problems in a wide range of disciplines.


2021 ◽  
Author(s):  
Francesco Padovani ◽  
Benedikt Mairhoermann ◽  
Pascal Falter-Braun ◽  
Jette Lengefeld ◽  
Kurt M Schmoller

Live-cell imaging is a powerful tool to study dynamic cellular processes on the level of single cells with quantitative detail. Microfluidics enables parallel high-throughput imaging, creating a downstream bottleneck at the stage of data analysis. Recent progress on deep learning image analysis dramatically improved cell segmentation and tracking. Nevertheless, manual data validation and correction is typically still required and broadly used tools spanning the complete range of live-cell imaging analysis, from cell segmentation to pedigree analysis and signal quantification, are still needed. Here, we present Cell-ACDC, a user-friendly graphical user-interface (GUI)-based framework written in Python, for segmentation, tracking and cell cycle annotation. We included two state-of-the-art and high-accuracy deep learning models for single-cell segmentation of yeast and mammalian cells implemented in the most used deep learning frameworks TensorFlow and PyTorch. Additionally, we developed and implemented a cell tracking method and embedded it into an intuitive, semi-automated workflow for label-free cell cycle annotation of single cells. The open-source and modularized nature of Cell-ACDC will enable simple and fast integration of new deep learning-based and traditional methods for cell segmentation or downstream image analysis. Source code: https://github.com/SchmollerLab/Cell_ACDC


2020 ◽  
Author(s):  
Young Hwan Chang ◽  
Jeremy Linsley ◽  
Josh Lamstein ◽  
Jaslin Kalra ◽  
Irina Epstein ◽  
...  

AbstractLive-cell imaging is an important technique to study cell migration and proliferation as well as image-based profiling of drug perturbations over time. To gain biological insights from live-cell imaging data, it is necessary to identify individual cells, follow them over time and extract quantitative information. However, since often biological experiment does not allow the high temporal resolution to reduce excessive levels of illumination or minimize unnecessary oversampling to monitor long-term dynamics, it is still a challenging task to obtain good tracking results with coarsely sampled imaging data. To address this problem, we consider cell tracking problem as “stable matching problem” and propose a robust tracking method based on Voronoi partition which adapts parameters that need to be set according to the spatio-temporal characteristics of live cell imaging data such as cell population and migration. We demonstrate the performance improvement provided by the proposed method using numerical simulations and compare its performance with proximity-based tracking and nearest neighbor-based tracking.


Author(s):  
James A. Cornwell ◽  
Maria Z. Gutierrez ◽  
Richard P. Harvey ◽  
Robert E. Nordon

2019 ◽  
Author(s):  
Vardan Andriasyan ◽  
Artur Yakimovich ◽  
Fanny Georgi ◽  
Anthony Petkidis ◽  
Robert Witte ◽  
...  

Imaging across scales gives insight into disease mechanisms in organisms, tissues and cells. Yet, rare infection phenotypes, such as virus-induced cell lysis have remained difficult to study. Here, we developed fixed and live cell imaging modalities and a deep learning approach to identify herpesvirus and adenovirus infections in the absence of virus-specific stainings. Procedures comprises staining of infected nuclei with DNA-dyes, fluorescence microscopy, and validation by virus-specific live-cell imaging. Deep learning of multi-round infection phenotypes identified hallmarks of adenovirus-infected cell nuclei. At an accuracy of >95%, the procedure predicts two distinct infection outcomes 20 hours prior to lysis, nonlytic (nonspreading) and lytic (spreading) infections. Phenotypic prediction and live-cell imaging revealed a faster enrichment of GFP-tagged virion proteins in lytic compared to nonlytic infected nuclei, and distinct mechanics of lytic and nonlytic nuclei upon laser-induced ruptures. The results unleash the power of deep learning based prediction in unraveling rare infection phenotypes.


2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Christian Carsten Sachs ◽  
Joachim Koepff ◽  
Wolfgang Wiechert ◽  
Alexander Grünberger ◽  
Katharina Nöh

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