RT Journal Article T1 A Noninvasive Method for Time-Lapse Imaging of Microbial Interactions and Colony Dynamics. A1 Molina-Santiago, Carlos A1 Pearson, John R A1 Berlanga-Clavero, María Victoria A1 Pérez-Lorente, Alicia Isabel A1 de Vicente, Antonio A1 Romero, Diego K1 biofilms K1 microbe-microbe interactions K1 microbial communities AB Complex interactions between microbial populations can greatly affect the overall properties of a microbial community, sometimes leading to cooperation and mutually beneficial coexistence, or competition and the death or displacement of organisms or subpopulations. Interactions between different biofilm populations are highly relevant in diverse scientific areas, from antimicrobial resistance to microbial ecology. The utilization of modern microscopic techniques has provided a new and interesting insight into how bacteria interact at the cellular level to form and maintain microbial biofilms. However, our ability to follow complex intraspecies and interspecies interactions in vivo at the microscopic level has remained somewhat limited. Here, we detailed BacLive, a novel noninvasive method for tracking bacterial growth and biofilm dynamics using high-resolution fluorescence microscopy and an associated ImageJ processing macro (https://github.com/BacLive) for easier data handling and image analysis. Finally, we provided examples of how BacLive can be used in the analysis of complex bacterial communities. IMPORTANCE Communication and interactions between single cells are continuously defining the structure and composition of microbial communities temporally and spatially. Methods routinely used to study these communities at the cellular level rely on sample manipulation which makes microscopic time-lapse experiments impossible. BacLive was conceived as a method for the noninvasive study of the formation and development of bacterial communities, such as biofilms, and the formation dynamics of specialized subpopulations in time-lapse experiments at a colony level. In addition, we developed a tool to simplify the processing and analysis of the data generated by this method. YR 2022 FD 2022-07-05 LK http://hdl.handle.net/10668/20037 UL http://hdl.handle.net/10668/20037 LA en DS RISalud RD Apr 6, 2025