Forming a Large-Scale Droplet Array in a Microcage Array Chip for High-Throughput Screening.,Analytical Chemistry

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Forming a Large-Scale Droplet Array in a Microcage Array Chip for High-Throughput Screening.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2019-07-23 00:00:00 , DOI: 10.1021/acs.analchem.9b02288
Jin-Gang Xu ₁ , Meng-Shi Huang ₁ , Hui-Feng Wang ₁ , Qun Fang ₁

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Forming a large-scale droplet array plays an important role for microfluidic droplet-based high-throughput screening and analysis. Herein, we describe a simple and rapid method to form a large-scale two-dimension (2D) droplet array by using a microcage array chip. Differing from the previous droplet array formation methods, microcages formed by being surrounded by multiple micropillars could rapidly spread the oil phase through the gaps between the micropillars and trap droplets with fast speed and convenient operation. We formed a large-scale 2D monolayer droplet array containing approximately 1 000 000 droplets on a 5.5 cm × 5.5 cm microcage array chip within 90 s. The droplets in the droplet array could be further incubated for performing biochemical reactions and detected by a fluorescence microscope in real time. Due to the exact trapping and positioning functions of the microcages to the droplets, single targeted fluorescent droplets in the array could be individually picked out and transferred to culture medium by a microfluidic droplet-handling robot with a success rate of 100% and a picking operation time of 2.0 s for one droplet under the optimized conditions. This system was validated in the screening of the bacterium expressing the esterase AFEST from a mixture of AFEST-expressing and phosphotriesterase-expressing E. coli cells, achieving a success rate of 100% for single-droplet picking while maintaining the bacterial cell viability. The present system has the potential to be applied in high-throughput screening and analysis, such as single cell analysis, directed evolution, and drug screening.

中文翻译：

在微笼阵列芯片中形成大规模液滴阵列，以进行高通量筛选。

形成大规模液滴阵列对于基于微流体液滴的高通量筛选和分析起着重要作用。在这里，我们描述了一种通过使用微笼阵列芯片来形成大规模二维（2D）液滴阵列的简单而快速的方法。与先前的液滴阵列形成方法不同，由多个微柱包围形成的微笼可以使油相通过微柱之间的间隙快速扩散，并以快速且方便的操作捕获液滴。在90 s内，我们在5.5 cm x 5.5 cm微笼阵列芯片上形成了一个包含约1000000液滴的大规模2D单层液滴阵列。液滴阵列中的液滴可以进一步孵育以进行生化反应，并通过荧光显微镜实时检测。由于微笼对液滴的精确捕获和定位功能，阵列中的单个目标荧光液滴可以通过微流体液滴处理机器人单独拾取并转移到培养基中，成功率达100％，并具有拾取操作在优化条件下，一滴液滴的时间为2.0 s。该系统在从表达AFEST和磷酸三酯酶的混合物中筛选出表达酯酶AFEST的细菌时得到了验证大肠杆菌细胞，在保持细菌细胞活力的同时，单滴采摘成功率达到100％。本系统具有潜力用于高通量筛选和分析，例如单细胞分析，定向进化和药物筛选。

更新日期：2019-07-23

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