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Microscopy image analysis is a major bottleneck in quantification of single-cell microscopy data, typically requiring human oversight and curation, which limit both accuracy and throughput. To address this, we developed a deep learning-based image analysis pipeline that performs segmentation, tracking, and lineage reconstruction. Our analysis focuses on time-lapse movies of  Escherichia coli  cells trapped in a "mother machine" microfluidic device, a scalable platform for long-term single-cell analysis that is widely used in the field. While deep learning has been applied to cell segmentation problems before, our approach is fundamentally innovative in that it also uses machine learning to perform cell tracking and lineage reconstruction. With this framework we are able to get high fidelity results (1% error rate), without human intervention. Further, the algorithm is fast, with complete analysis of a typical frame containing ~150 cells taking <700msec. The framework is not constrained to a particular experimental set up and has the potential to generalize to time-lapse images of other organisms or different experimental configurations. These advances open the door to a myriad of applications including real-time tracking of gene expression and high throughput analysis of strain libraries at single-cell resolution.

DeLTA: Automated cell segmentation, tracking, and lineage reconstruction using deep learning