Exploring the emergence of single-cell heterogeneity in a developing biofilm

Biofilms consist of surface-associated bacteria embedded in an extracellular polymeric matrix. This matrix provides protection and allows for resource exchange within the community. Although bacterial colonies are known to exhibit broad levels of heterogeneity, it is not well understood how this extends to biofilms. Recently, we’ve found multiple heterogeneities on developing biofilms grown from a single founding cell. These heterogeneities are coupled to the cells’ mechanical environment and their structural organization, exemplifying how variations on biofilm regulation, cell organization and local mechanical environments are correlated in the biofilm development process. How these heterogeneities emerge in the developing biofilm, and their effect on the biofilm’s lifecycle remains largely unknown. Here, we use the model organism Vibrio cholerae to study the emergence of spatiotemporal patterning and heterogeneity within biofilms at the single-cell level. We combine high-resolution time-lapse imaging, fluorescent reporters and single-cell tracking to perform lineage tracing in a developing biofilm. With this novel technique, we aim to study non-genetic inheritance in the biofilm and its consequences on role division and function.