Automated deep lineage tree analysis using a Bayesian single cell tracking approach

Automated deep lineage tree analysis using a Bayesian single cell tracking approach
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使用贝叶斯单细胞跟踪方法进行自动深度谱系树分析

DOI:
10.1101/2020.09.10.276980
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发表时间:
2020
期刊:
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通讯作者:
Ulicna K
Ulicna K
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作者:
Ulicna K

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单细胞方法开始揭示细胞群体的内在异质性,这是由确定性和随机性过程的相互作用引起的。然而,从延时显微镜数据中量化单细胞行为仍然具有挑战性,因为难以在长时间尺度和几代人之间提取可靠的细胞轨迹和谱系信息。因此,我们开发了一种混合深度学习和贝叶斯细胞跟踪方法,以从活细胞显微镜数据中重建谱系树。我们实现了一个残差U-Net模型与分类CNN相结合,以实现细胞核的准确实例分割。为了随着时间的推移和通过细胞分裂来跟踪细胞,我们开发了一种贝叶斯细胞跟踪方法,该方法使用来自图像的输入特征来从数千小时的活细胞成像数据库中检索多代谱系信息。使用我们的方法,我们从超过3,500小时的视频片段中提取了20,000多个完全注释的单细胞轨迹,组织成跨越八代和第四代表亲距离的多代谱系树。基准测试,包括谱系树重建评估,表明我们的方法产生高保真度的结果与我们的数据,手动策展的要求最低。为了证明我们的最低监督细胞跟踪方法的鲁棒性,我们在5,000多个完全注释的细胞谱系中检索细胞周期持续时间及其扩展的代际和代内家族关系。我们观察到消失的周期持续时间的相关性在祖先的亲戚,但揭示相关的细胞之间共享同一代的扩展谱系。这些发现扩展了所研究的细胞谱系关系的深度和广度,比以前依赖于半手动谱系数据分析的细胞周期遗传性研究多了大约两个数量级的数据。
Single-cell methods are beginning to reveal the intrinsic heterogeneity in cell populations, arising from the interplay of deterministic and stochastic processes. However, it remains challenging to quantify single-cell behaviour from time-lapse microscopy data, owing to the difficulty of extracting reliable cell trajectories and lineage information over long time-scales and across several generations. Therefore, we developed a hybrid deep learning and Bayesian cell tracking approach to reconstruct lineage trees from live-cell microscopy data. We implemented a residual U-Net model coupled with a classification CNN to allow accurate instance segmentation of the cell nuclei. To track the cells over time and through cell divisions, we developed a Bayesian cell tracking methodology that uses input features from the images to enable the retrieval of multi-generational lineage information from a corpus of thousands of hours of live-cell imaging data. Using our approach, we extracted 20,000 + fully annotated single-cell trajectories from over 3,500 h of video footage, organised into multi-generational lineage trees spanning up to eight generations and fourth cousin distances. Benchmarking tests, including lineage tree reconstruction assessments, demonstrate that our approach yields high-fidelity results with our data, with minimal requirement for manual curation. To demonstrate the robustness of our minimally supervised cell tracking methodology, we retrieve cell cycle durations and their extended inter- and intra-generational family relationships in 5,000 + fully annotated cell lineages. We observe vanishing cycle duration correlations across ancestral relatives, yet reveal correlated cyclings between cells sharing the same generation in extended lineages. These findings expand the depth and breadth of investigated cell lineage relationships in approximately two orders of magnitude more data than in previous studies of cell cycle heritability, which were reliant on semi-manual lineage data analysis.
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文章标题:从低时间分辨率的高通量显微镜中提取细胞谱系的荧光报告基因时间过程从低时间分辨率的高通量显微镜中提取细胞谱系的荧光报告基因时间过程
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