A tool named Iris for versatile high-throughput phenotyping in microorganisms.

Abstract

Advances in our ability to systematically introduce and track controlled genetic variance in microbes have fueled high-throughput reverse genetics approaches in the past decade. When coupled to quantitative readouts, such approaches are extremely powerful at elucidating gene function and providing insights into the underlying pathways and the overall cellular network organization. Yet, until now all efforts for quantifying microbial macroscopic phenotypes have been restricted to monitoring growth in a small number of model microbes. We developed an image analysis software named Iris, which allows for systematic exploration of a number of orthogonal-to-growth processes, including biofilm formation, colony morphogenesis, envelope biogenesis, sporulation and reporter activity. In addition, Iris provides more sensitive growth measurements than current available software, and is compatible with a variety of different microbes, as well as with endpoint or kinetic data. We used Iris to reanalyze existing chemical genomics data in Escherichia coli and to perform proof-of-principle screens on colony biofilm formation and morphogenesis of different bacterial species and the pathogenic fungus, Candida albicans. Thereby we recapitulated existing knowledge but also identified a plethora of additional genes and pathways involved in both processes.