Department of Informatics, Technical University of Munich, Germany; Institute for Advanced Study, Technical University of Munich, Germany.
Faculty of Physics, Ludwig-Maximilians University of Munich, Germany.
Med Image Anal. 2018 Aug;48:147-161. doi: 10.1016/j.media.2018.05.009. Epub 2018 Jun 5.
In vitro experiments with cultured cells are essential for studying their growth and migration pattern and thus, for gaining a better understanding of cancer progression and its treatment. Recent progress in lens-free microscopy (LFM) has rendered it an inexpensive tool for label-free, continuous live cell imaging, yet there is only little work on analysing such time-lapse image sequences. We propose (1) a cell detector for LFM images based on fully convolutional networks and residual learning, and (2) a probabilistic model based on moral lineage tracing that explicitly handles multiple detections and temporal successor hypotheses by clustering and tracking simultaneously. (3) We benchmark our method in terms of detection and tracking scores on a dataset of three annotated sequences of several hours of LFM, where we demonstrate our method to produce high quality lineages. (4) We evaluate its performance on a somewhat more challenging problem: estimating cell lineages from the LFM sequence as would be possible from a corresponding fluorescence microscopy sequence. We present experiments on 16 LFM sequences for which we acquired fluorescence microscopy in parallel and generated annotations from them. Finally, (5) we showcase our methods effectiveness for quantifying cell dynamics in an experiment with skin cancer cells.
体外细胞培养实验对于研究细胞的生长和迁移模式至关重要,有助于深入了解癌症的进展及其治疗方法。无透镜显微镜(LFM)的最新进展使其成为一种用于无标记、连续活细胞成像的廉价工具,但对这种时程图像序列的分析研究还很少。我们提出了(1)一种基于全卷积网络和残差学习的 LFM 图像细胞检测方法,以及(2)一种基于道德谱系追踪的概率模型,通过聚类和跟踪同时显式处理多个检测和时间后继假设。(3)我们在三个经过注释的数小时 LFM 图像序列数据集上,根据检测和跟踪分数对我们的方法进行基准测试,结果表明我们的方法能够生成高质量的谱系。(4)我们还评估了该方法在更具挑战性的问题上的性能:从 LFM 序列中估计细胞谱系,就像从相应的荧光显微镜序列中获得一样。我们在 16 个 LFM 序列上进行了实验,我们从这些序列中获取了荧光显微镜图像,并从中生成了注释。最后,(5)我们展示了我们的方法在皮肤癌细胞实验中定量分析细胞动力学的有效性。