As a reference for unmodified division rings, we imaged Z-rings in E. coli cells expressing FtsZ-mNeonGreen as the only source of FtsZ22. Under our experimental conditions, this strain produced normal-looking, sharp Z-rings (Supplementary Figure 1) and grew and divided similarly to wild-type (WT) E. coli (MC4100) (Supplementary Figure 2a-e). We then trapped the cells in a vertical position in micron-sized holes that were produced in agarose pads using silica micron pillar arrays14 (Fig.