4D monitoring of bacterial biofilm growth at the air-liquid interface with optical coherence interference

While bacteria are mostly found within bacterial biofilms in nature, the physical mechanisms that support biofilm formation at the air-liquid interface (ALI) are often elusive, in part due to the lack of quantitative imaging methods. Here, we introduce a novel setup integrating Spectral Domain Optical Coherence Tomography (SD-OCT) in a custom-built incubator for non-invasive, longitudinal, label-free monitoring of bacterial biofilms growth at the ALI. This approach allowed for high-resolution quantification of biofilm 3D structure within a 2.5×2.5×3mm³ field of view continuously for 40 hours. Although this proceeding focuses on the experimental procedure as well as the automated analysis pipeline, this setup is currently being used to study the physicochemical and genetic factors that drive biofilms formation.