Fig. 3
Flow cytometric analysis of sorting UPWRP_1 from an activated sludge sample hybridised with probes Ribo_Unk1029_17Cy5 and EUB388A488. An ~100,000 events were collected for each FACS plot, except for the purity check. Sorting gates are outlined in black and values shown indicate the percentage of gated events over the total number of events. Flow cytometric analysis: a Sorting gate 1: sorting of bacterial cells using their light-scattering properties with a plot of forward versus side scatter; b sorting gate 2: filtering out cell aggregates based on side scatter area versus side scatter height; c sorting gate 3: filtering out cell aggregates based on forward scatter area versus forward scatter height. Sorting gate 4 was constructed to exclude events exhibiting Cy5 and A488 fluorescence signal in the negative controls: d no-probe control; e hybridisation with probe NON338Cy5 to estimate non-specific binding for Cy5 fluorophore; f hybridisation with probe NON338A488 to estimate non-specific binding for A488 fluorophore. g Events exhibiting Cy5 and A488 fluorescence signal above the cut-off threshold for the negative controls were collected in sorting gate 4. h Purity of the sorted sample after an initial round of sorting
