Rapid screening of antibiotic toxicity in an automated microdroplet system
Rapid screening of antibiotic toxicity in an automated microdroplet system
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DOI：
10.1039/c2lc21284f
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发表时间：
2012-01-01
期刊：
影响因子：
6.1
通讯作者：
Garstecki, Piotr
中科院分区：
文献类型：
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作者：
Churski, Krzysztof;Kaminski, Tomasz S.;Garstecki, Piotr
我们报告了一个自动化的微流控平台，用于“数字化”筛选含有小分子鸡尾酒的液滴的组成空间，并通过研究抗生素和大肠杆菌ATCC 25922之间的上位相互作用来展示该系统的功能。该系统具有几个关键特征：（i）它使用小型（ii）它产生具有组合物的液滴序列，所述组合物包括由用户编程的试剂和细菌细胞悬浮液;（iii）其将液滴序列输出到管的外部段，该管的外部段随后被断开以用于孵育和储存;和（iv）在培养液滴中的细菌后，将液滴注入配备有测量细胞生长的在线光纤分光光度计的第二装置中。该系统生成并融合液滴，并精确控制液体体积和输入底物的浓度（标准偏差< 1%）。我们展示了该技术在确定氨苄青霉素、四环素和氯霉素对大肠杆菌的最小抑菌浓度和成对相互作用中的应用。杆菌这些实验消耗少量试剂，需要几分钟来产生液滴，并需要几个小时进行孵育和分析。
We report an automated microfluidic platform for 'digitally' screening the composition space of droplets containing cocktails of small molecules and demonstrate the features of this system by studying epistatic interactions between antibiotics and Escherichia coli ATCC 25922. This system has several key characteristics: (i) it uses small (< 100 mu L) samples of liquids and suspensions of bacteria that are introduced directly into the chip; (ii) it generates a sequence of droplets with compositions, including reagents and bacterial cell suspensions that are programmed by the user; (iii) it exports the sequence of droplets to an external segment of tubing that is subsequently disconnected for incubation and storage; and (iv) after incubation of bacteria in droplets, the droplets are injected into a second device equipped with an in-line fiber optic spectrophotometer that measures cell growth. The system generates and fuses droplets with precise (< 1% in standard deviation) control of liquid volumes and of the concentrations of input substrates. We demonstrate the application of this technology in determining the minimum inhibitory concentration and pair-wise interactions of ampicillin, tetracycline, and chloramphenicol against E. coli. The experiments consumed small volumes of reagents and required minutes to create the droplets and several hours for their incubation and analysis.