Analytical Sciences

A Bacterial Continuous Culture System Based on a Microfluidic Droplet Open Reactor

Manami ITO,*1 Haruka SUGIURA,*1 Shotaro AYUKAWA,*2 Daisuke KIGA,*1,*3 and Masahiro TAKINOUE*1,*4

*1 Department of Computational Intelligence and Systems Science, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori, Yokohama, Kanagawa 226-8502, Japan
*2 Education Academy of Computational Life Sciences (ACLS), Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori, Yokohama, Kanagawa 226-8502, Japan
*3 Earth-Life Science Institute (ELSI), Tokyo Institute of Technology, 2-12-1 Ookayama, Meguro, Tokyo 152-8550, Japan
*4 PRESTO, Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan

Recently, micrometer-sized bacterial culture systems have attracted attention as useful tools for synthetic biology studies. Here, we present the development of a bacterial continuous culture system based on a microdroplet open reactor consisting of two types of water-in-oil microdroplets with diameters of several hundred micrometers. A continuous culture was realized the through supply of nutrient substrates and the removal of waste and excess bacterial cells based on repeated fusion and fission of droplets. The growth dynamics was controlled by the interval of fusion. We constructed a microfluidic system and quantitatively assessed the dynamics of the bacterial growth using a mathematical model. This system will facilitate the study of synthetic biology and metabolic engineering in the future.

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