Dynamical analysis of bacteria in microscopy movies

PLoS One. 2019 Jun 6;14(6):e0217823. doi: 10.1371/journal.pone.0217823. eCollection 2019.

Abstract

Recent advances in microscopy, computing power and image processing have enabled the analysis of ever larger datasets of movies of microorganisms to study their behaviour. However, techniques for analysing the dynamics of individual cells from such datasets are not yet widely available in the public domain. We recently demonstrated significant phenotypic heterogeneity in the adhesion of Escherichia coli bacteria to glass surfaces using a new method for the high-throughput analysis of video microscopy data. Here, we present an in-depth analysis of this method and its limitations, and make public our algorithms for following the positions and orientations of individual rod-shaped bacteria from time-series of 2D images to reconstruct their trajectories and characterise their dynamics. We demonstrate in detail how to use these algorithms to identify different types of adhesive dynamics within a clonal population of bacteria sedimenting onto a surface. The effects of measurement errors in cell positions and of limited trajectory durations on our results are discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Bacterial Adhesion
  • Diffusion
  • Escherichia coli / cytology*
  • Microscopy, Video*
  • Reproducibility of Results
  • Rotation
  • Surface Properties

Grants and funding

Funding was provided through FP7-PEOPLE-2013-IEF Marie Curie fellowship LivPaC, 623364 (TV), Marie Curie fellowship H2020-MSCA-IF-2014 ActiDoC, 654688 (NK), ERC Advanced Grant GA 340877-PHYSAPS (TV, NK) and an EPSRC programme grant EP/J007404/1 (AD, WP, JSL). JSL was also funded by the National Physical Laboratory and SUPA. ATB received funding from UK EPSRC (EP/S001255/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.