Here we describe a procedure to make bacterial communication assays based on 3OC6HSL and 3OC12HSL quorum sensing signaling. Strains that sense and response to these signals are plated on a permeable membrane printed with a hydrophobic grid, which is placed on top of an agar plate containing growth media and antibiotics. This method is based on Grant et al. 2016 (Grant PK, Dalchau N, Brown JR, Federici F, Rudge TJ, Yordanov B, Patange O, Phillips A, Haseloff J.Orthogonal Intercellular Signaling for Programmed Spatial Behavior. Mol Syst Biol. 2016 Jan 25;12(1):849. doi: 10.15252/msb.20156590)This assay has been used for low cost and open source fluorescence imaging (please see https://osf.io/dy6p2/ for further information & data, https://github.com/SynBioUC/FluoPi for code and http://docubricks.com/viewer.jsp? for hardware assembly)