Extending incubation to 16 d enhances the formation of PAO1 BLS. Bacterial inoculation and incubation for the development of BLS were done as described in Figure 1, except fresh ASM+ was added to the wells at 4-d intervals to replace lost volume. (A) CLSM micrographs of BLS at 16 d post-inoculation; magnification, 10X; bar, 200.00 nm. (B) The 3-D architecture of the BLS shown in (A). Boxes, 800.00 px W x 600.00 px H; bar, 100 px.