A Microfluidic Platform for High-Throughput Screening of Small Mutant Libraries.
The screening and isolation of target microorganisms from mutated recombinant libraries are crucial for the advancement of synthetic biology and metabolic engineering. However, conventional screening tools present several limitations in throughput, cost, and labor. Herein, we describe a novel microfluidic high-throughput screening (HTS) platform with several advantages. The platform utilizes a fluid array to compartmentalize bacterial cells in well-ordered separated microwells and allows long-term cell culture with high throughput. The platform enables the extraction of selected target cells from the fluid array for additional culture and postanalysis by using a capillary-driven sample relocation method. To confirm the feasibility of the platform, we demonstrated two different types of HTS methods based on the levels of reporter gene expression and cellular growth rate difference. For the reporter gene-based HTS, a spike recovery approach was taken to demonstrate that target cells are successfully screened out from a mixture containing nontarget cells by repeating the culture and extraction processes. Additionally, the same platform allowed us to screen and sort target cells according to their cellular growth rate difference, which seems hard in conventional screening methods. Hence, the platform could be used for various microbiological assays, including the detection of cell-excreted metabolites, microbial biosensors, and other HTS systems.
Department of Biomedical Engineering, ‡Department of Mechanical Engineering, and §Department of Chemical Engineering, Ulsan National Institute of Science and Technology , 50 UNIST-gil, Ulsan 44919, Republic of Korea.
Ji Won
JW
Lim
Department of Biomedical Engineering, ‡Department of Mechanical Engineering, and §Department of Chemical Engineering, Ulsan National Institute of Science and Technology , 50 UNIST-gil, Ulsan 44919, Republic of Korea.
Kwang Soo
KS
Shin
Department of Biomedical Engineering, ‡Department of Mechanical Engineering, and §Department of Chemical Engineering, Ulsan National Institute of Science and Technology , 50 UNIST-gil, Ulsan 44919, Republic of Korea.
Jaemin
J
Moon
Department of Biomedical Engineering, ‡Department of Mechanical Engineering, and §Department of Chemical Engineering, Ulsan National Institute of Science and Technology , 50 UNIST-gil, Ulsan 44919, Republic of Korea.
Sung Kuk
SK
Lee
Department of Biomedical Engineering, ‡Department of Mechanical Engineering, and §Department of Chemical Engineering, Ulsan National Institute of Science and Technology , 50 UNIST-gil, Ulsan 44919, Republic of Korea.
Taesung
T
Kim
D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays; D046228:Microarray Analysis; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000494:pharmacology
D054852:Small Molecule Libraries