For initial experiments, microfluidic devices were fabricated from an epoxy replica of the mother machine described by Potvin-Trottier et al. (24) (fig. S1, design A). The replica mold was a gift from M. Cabeen (Harvard University). Subsequently, custom SU-8 molds were generated to build channels that better matched the typical cell width in our growth conditions (fig. S1, design B). See the “Microfluidic master fabrication” section in the Supplementary Materials for details.

To prepare a new microfluidic device out of the mold, dimethylsiloxane monomer (Sylgard 184, Dow Corning) was mixed with the curing agent at a 10:1 ratio, degassed, and poured over the mold. This mixture was degassed for an additional 1 hour and cured overnight at 65°C before being removed from the mold. Individual devices were first cut from the cured polydimethylsiloxane. Subsequently, inlets and outlets for each flow channel (“lane”) were created using a 0.75-mm biopsy punch. The device was treated with oxygen plasma in a plasma cleaner along with a 25 mm × 40 mm No. 1.5 coverglass (VWR) for 15 s at 30 W and an oxygen pressure of 200 mtorr. Following plasma treatment, the device and glass were bonded and incubated at least 1 hour at 65°C before use.

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