Double-Sided Microwells with a Stepped Through-Hole Membrane for High-Throughput Microbial Assays
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- Double-Sided Microwells with a Stepped Through-Hole Membrane for High-Throughput Microbial Assays
- Bae, Juyeol; Ju, Janghyun; Kim, Dahyun; Kim, Taesung
- Issue Date
- AMER CHEMICAL SOC
- ANALYTICAL CHEMISTRY, v.92, no.14, pp.9501 - 9510
- To improve the throughput of microwell arrays for identifying immense cellular diversities even at a single-bacteria level, further miniaturization or densification of the microwells has been an obvious breakthrough. However, controlling millions of nanoliter samples or more at the microscale remains technologically difficult and has been spatially restricted to a single open side of the microwells. Here we employed a stepped through-hole membrane to utilize the bottom as well as top side of a high-density nanoliter microwell array, thus improving spatial efficiency. The stepped structure shows additional effectiveness for handling several millions of nanoliter bacterial samples in the overall perspectives of controllability, throughput, simplicity, versatility, and automation by using novel methods for three representative procedures in bacterial assays: partitioning cells, manipulating the chemical environment, and extracting selected cells. As a potential application, we show proof-of-concept isolation of rare cells in a mixed ratio of 1 to around 10(6) using a single chip. Our device can be further applied to various biological studies pertaining to synthetic biology, drug screening, mutagenesis, and single-cell heterogeneity.
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