Optogenetics reprogramming of planktonic cells for biofilm formation.
                            
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                                BphS
                            
                            
                                
                                    P. aeruginosa
                                
                            
                            
                                Control of cytoskeleton / cell motility / cell shape
                            
                                Control of cell-cell / cell-material interactions
                            
                                Immediate control of second messengers
                            
                            
                            
                            
                            
                        
                        
                        
                        
                            Abstract:
                            Single-cell behaviors play essential roles during early-stage biofilms formation. In this study, we evaluated whether biofilm formation could be guided by precisely manipulating single cells behaviors. Thus, we established an illumination method to precisely manipulate the type IV pili (TFP) mediated motility and microcolony formation of Pseudomonas aeruginosa by using a combination of a high-throughput bacterial tracking algorithm, optogenetic manipulation and adaptive microscopy. We termed this method as Adaptive Tracking Illumination (ATI). We reported that ATI enables the precise manipulation of TFP mediated motility and microcolony formation during biofilm formation by manipulating bis-(3′-5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) levels in single cells. Moreover, we showed that the spatial organization of single cells in mature biofilms can be controlled using ATI. Thus, the established method (i.e., ATI) can markedly promote ongoing studies of biofilms.