Fig. 1.
High-throughput time-resolved imaging methodology. Shown is a schematic representation of the high-throughput time-resolved imaging methodology. Strains are grown in glass-bottom 96-well plates and a perturbant is added to the desired wells. Morphological changes are monitored over time by automated phase contrast imaging. Image acquisition is dynamically regulated by automatically defining the exposure and number of images to be collected for every well depending on the cell density and cell morphology. Four or nine images were collected per well. The imaging protocol yields a rich dataset providing time-resolved records of phenotypes. In this work, 4218 strains from the Keio collection of non-essential single-gene deletion mutants of E. coli were perturbed by the antibiotic cefsulodin and images were recorded at three time-points leading to a dataset including approximately 60,000 frames and 2 million single cells. The image dataset is then subjected to image and data processing. Scale bar corresponds to 20 μm