Microfluidic Device for Automated Synchronization of Bacterial Cells

Abstract

We report the development of an automated microfluidic “baby machine” to synchronize the bacterium <i>Caulobacter crescentus</i> on-chip and to move the synchronized populations downstream for analysis. The microfluidic device is fabricated from three layers of poly­(dimethylsiloxane) and has integrated pumps and valves to control the movement of cells and media. This synchronization method decreases incubation time and media consumption and improves synchrony quality compared to the conventional plate-release technique. Synchronized populations are collected from the device at intervals as short as 10 min and at any time over four days. Flow cytometry and fluorescence cell tracking are used to determine synchrony quality, and cell populations synchronized in minimal growth medium with 0.2% glucose (M2G) and peptone yeast extract (PYE) medium contain >70% and >80% swarmer cells, respectively. Our on-chip method overcomes limitations with conventional physical separation methods that consume large volumes of media, require manual manipulations, have lengthy incubation times, are limited to one collection, and lack precise temporal control of collection times