A programmable droplet-based microfluidic device applied to multiparameter analysis of single microbes and microbial communities
A programmable droplet-based microfluidic device applied to multiparameter analysis of single microbes and microbial communities
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DOI：
10.1073/pnas.1106752109
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发表时间：
2012-05-15
影响因子：
11.1
通讯作者：
Hansen, Carl L.
中科院分区：
文献类型：
--
作者：
Leung, Kaston;Zahn, Hans;Hansen, Carl L.
我们提出了一种可编程的基于液滴的微流体装置，其结合了集成微阀技术的可重构流动路由能力与液滴固有的样品区室化和无分散运输。该设备允许通过连续合并含有试剂或细胞的皮升体积液滴的可编程序列，在95个可单独寻址的纳升体积储存室中执行用户定义的多步反应方案。该功能通过“流控润湿”来实现，“流控润湿”是一种液滴对接和合并机制，其利用通过通道的液滴流的物理特性来控制液滴润湿的精确位置。该设备还允许自动无交叉污染地将反应产物从各个腔室回收到标准微量离心管中用于下游分析。可编程性、可寻址性和选择性恢复的组合特征提供了可针对多种应用重新编程的通用硬件平台。我们通过使用该设备实施多个单细胞实验类型来证明这种多功能性：细菌细胞分选和培养，分类基因鉴定，以及使用常见实验室菌株的高通量单细胞全基因组扩增和测序。最后，我们将该设备应用于来自不同环境样本的单细胞和微生物财团的基因组分析，包括海洋富集培养物，深海沉积物和人类口腔。由此产生的数据集捕获单个细胞的基因型特性，并阐明微生物群落成员之间已知的和潜在的独特伙伴关系。
We present a programmable droplet-based microfluidic device that combines the reconfigurable flow-routing capabilities of integrated microvalve technology with the sample compartmentalization and dispersion-free transport that is inherent to droplets. The device allows for the execution of user-defined multistep reaction protocols in 95 individually addressable nanoliter-volume storage chambers by consecutively merging programmable sequences of picoliter-volume droplets containing reagents or cells. This functionality is enabled by "flow-controlled wetting," a droplet docking and merging mechanism that exploits the physics of droplet flow through a channel to control the precise location of droplet wetting. The device also allows for automated cross-contamination-free recovery of reaction products from individual chambers into standard microfuge tubes for downstream analysis. The combined features of programmability, addressability, and selective recovery provide a general hardware platform that can be reprogrammed for multiple applications. We demonstrate this versatility by implementing multiple single-cell experiment types with this device: bacterial cell sorting and cultivation, taxonomic gene identification, and high-throughput single-cell whole genome amplification and sequencing using common laboratory strains. Finally, we apply the device to genome analysis of single cells and microbial consortia from diverse environmental samples including a marine enrichment culture, deep-sea sediments, and the human oral cavity. The resulting datasets capture genotypic properties of individual cells and illuminate known and potentially unique partnerships between microbial community members.