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Image-Based Single Cell Profiling: High-Throughput Processing of Mother Machine Experiments

Christian Carsten Sachs, Alexander Grünberger, Stefan Helfrich, Christopher Probst, Wolfgang Wiechert, Dietrich Kohlheyer and Katharina Nöh

PLOS ONE, 2016, vol. 11, issue 9, 1-15

Abstract: Background: Microfluidic lab-on-chip technology combined with live-cell imaging has enabled the observation of single cells in their spatio-temporal context. The mother machine (MM) cultivation system is particularly attractive for the long-term investigation of rod-shaped bacteria since it facilitates continuous cultivation and observation of individual cells over many generations in a highly parallelized manner. To date, the lack of fully automated image analysis software limits the practical applicability of the MM as a phenotypic screening tool. Results: We present an image analysis pipeline for the automated processing of MM time lapse image stacks. The pipeline supports all analysis steps, i.e., image registration, orientation correction, channel/cell detection, cell tracking, and result visualization. Tailored algorithms account for the specialized MM layout to enable a robust automated analysis. Image data generated in a two-day growth study (≈ 90 GB) is analyzed in ≈ 30 min with negligible differences in growth rate between automated and manual evaluation quality. The proposed methods are implemented in the software molyso (MOther machine AnaLYsis SOftware) that provides a new profiling tool to analyze unbiasedly hitherto inaccessible large-scale MM image stacks. Conclusion: Presented is the software molyso, a ready-to-use open source software (BSD-licensed) for the unsupervised analysis of MM time-lapse image stacks. molyso source code and user manual are available at https://github.com/modsim/molyso.

Date: 2016
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Persistent link: https://EconPapers.repec.org/RePEc:plo:pone00:0163453

DOI: 10.1371/journal.pone.0163453

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