期刊
MICROFLUIDICS AND NANOFLUIDICS
卷 18, 期 3, 页码 493-501出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s10404-014-1447-2
关键词
Microfluidics; Concentration gradient; Droplet array; Protein crystallization; Screening
资金
- Natural Science Foundation of China [21305010, 21375012]
- Fundamental Research Funds for the Central Universities [N110805001, N110705002, N120305002]
- General Scientific Research Projects of Liaoning Provincial Department of Education [L2013106]
We developed a two-layer microfluidic chip for the generation of two-dimensional concentration-gradient droplet arrays. All of the channels, including a concentration-gradient formation channel network, an oil phase channel network, and droplet generators, were ingeniously arranged on a single polydimethylsiloxane (PDMS) substrate, which facilitates the fabrication of the chip. Ninety-six lines of subnanoliter droplets with concentration gradients can be formed on a 6-inlet PDMS chip. With the aqueous phase flow rate of 1.0 mu L/min and the oil phase flow rate of 35 mu L/min, relative standard deviation of the droplet sizes in the same channel was below 5.0 %, while in the 96 channels was 7.0 %. Multiparameter screening of protein crystallization conditions was demonstrated by using the two-dimensional concentration-gradient droplet arrays. Protein, precipitant, and buffer each at 16 concentrations for lysozyme crystallization were tested simultaneously in one 6-inlet chip, and five precipitants each at eight concentrations were screened at the same time in one 10-inlet chip. The concentration-gradient droplet chip proved to be a useful tool for screening crystallization conditions, characterized by low consumption, high throughput, convenience, and automatization.
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