Quorum sensing (QS) describes the capability of microbes to communicate with each other by the aid of small molecules. Here we investigate the dynamics of QS-regulated gene expression induced by acylhomoserine lactones (AHLs) in Pseudomonas putida IsoF containing a green fluorescent protein-based AHL reporter. The fluorescence time course of individual colonies is monitored following the external addition of a defined AHL concentration to cells which had previously reached the QS-inactive state in AHL-free medium. Using a microfluidic setup the experiment is performed both under flow and non-flow conditions. We find that without supplying external AHL gene expression is induced without flow while flow suppresses the induction. Both without and with flow, at a low AHL concentration the fluorescence onset is significantly delayed while fluorescence starts to increase directly upon the addition of AHL at a high concentration. The differences between no flow and flow can be accounted for using a two-compartment model. This indicates AHL accumulation in a volume which is not affected by the flow. The experiments furthermore show significant cell-to-cell and colony-to-colony variability which is discussed in the context of a compartmentalized QS mechanism.