A novel method for high-throughput data collection in predictive microbiology: Optical density monitoring of colony growth as a function of time

Recently, the focus of predictive food microbiology has shifted towards more mechanistically-inspired modelling. Together with this trend, the need for methods that allow rapid data collection at the (intra)cellular level, as well as the intermediate subpopulation/colony level, has emerged. Although several experimental techniques are currently available to study colony dynamics in/on solid media, their widespread implementation as high-throughput methods remains a challenge.In this research, a novel method is presented to study colony growth based on optical density measurements performed in microtiter plates. An area scan procedure was applied to monitor individual Escherichia coli colonies in 48-well plates at 30 °C. Based on a fixed threshold value to separate the object (colony) from the background, the colony area was determined as a function of time.With this technique, expansion of the colony in radial direction could be monitored. Practical limitations (i.e., maximum achievable resolution and colony size) of the proposed method were investigated. A comparison was made with existing methods at the level of hardware requirements, data acquisition and data processing. Overall, the novel optical density method proved to be a flexible, high-throughput tool for monitoring (the mechanisms of) microbial colony growth in solid(like) systems.

► A novel method was explored to study microbial behaviour in solid systems. ► Escherichia coli colonies were monitored with optical density measurements in microplates. ► Colony area was determined from a detailed area scan of the microplate well. ► Main limitations of the method are maximum resolution and speed of data acquisition. ► The method is flexible, rapid and a competitive alternative to existing methods.