Development of a microfluidic cell culture system for the adaptive laboratory evolution

Abstract

We construct a microfluidics-based long-term cell culture system that can perform multiple adaptive laboratory evolution (ALE) experiments in parallel under different conditions. Single E. coli cells were isolated into water droplets in oil (W/O emulsion) containing agarose gel. Micrometric gel balls (GBs) were then extracted from the oil phase and placed in a microfluidic perfusion culture device, which can generate a substrate concentration gradient. The growth of the clonal colonies was monitored under each condition. We tested this GB-based culture device to expand the application of ALE after the gene knockout based on the flux balance analysis (FBA) performed in our previous studies.