Chapter 2 - Time-lapse microscopy and image analysis of Escherichia coli cells in mother machines

Publication date: 2016Source: Methods in Microbiology, Volume 43Author(s): Y. Yang, X. Song, A.B. LindnerAbstractHigh-throughput longitudinal measurements of single cells by automated time-lapse microscopy, combined with dynamic system modelling, have in recent decades advanced our understanding of microbial physiology, such as cell size control, gene expression and regulation, subcellular organisation and organelle dynamics, development and cell fate decision. However, the sheer large numbers of cells resulting from exponential growth often limit both the experimental duration and numbers of single cells quantified. To address these issues, microfluidic devices are specifically designed to spatially track and align single cells or lineages of cells. In this chapter, we use the example of tracking Escherichia coli cells in mother machines to introduce methods and potential issues in the design, execution and image analysis of such experiments. For an experimental design, we highlight microfluidic and time-lapse microscopy considerations important for the successful interfacing between the two technologies and for obtaining easily quantifiable images. Then we provided typical protocols in executing such experiments. Finally, we devote a large portion of this chapter to introduce image analysis methods automating the segmentation, tracking and quantification of cell images obtained from mother machine experiments.
Source: Methods in Microbiology - Category: Microbiology Source Type: research