For time-lapse experiments, a passivating agent (Pluronic F-108, Sigma-Aldrich) was added to the growth medium at a final concentration of 0.85 g/liter. E. coli strains were first grown overnight in medium without the passivating agent. Cells were diluted the morning of the experiment 1:100 fold in fresh medium, containing the passivating agent, and allowed to grow to late exponential or early stationary phase. The reduced cell size at this growth phase improved the efficiency with which cells loaded in the device.

The cell culture was loaded into the inlet of the device by pipetting. The device was then centrifuged on a custom adaptor fit into a standard tabletop centrifuge at 6000g for 10 min. The inlets were connected to syringes filled with the growth medium (+passivating agent) via Tygon tubing (VWR; inside diameter 0.02 inch × outside diameter 0.06 inch). The flow-through was collected from the outlet via a second Tygon tubing into an empty beaker. The growth medium was first pumped at a rate of 35 μl/min for at least 1 hour to allow the inlets and outlets to be cleared. Afterward, the flow rate was reduced to 4 to 5 μl/min for the duration of the experiment.

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