Brouzes Eric, Medkova Martina, Savenelli Neal, Marran Dave, Twardowski Mariusz, Hutchison J Brian, Rothberg Jonathan M, Link Darren R, Perrimon Norbert, Samuels Michael L
RainDance Technologies, Lexington, MA 02421, USA.
Proc Natl Acad Sci U S A. 2009 Aug 25;106(34):14195-200. doi: 10.1073/pnas.0903542106. Epub 2009 Jul 15.
We present a droplet-based microfluidic technology that enables high-throughput screening of single mammalian cells. This integrated platform allows for the encapsulation of single cells and reagents in independent aqueous microdroplets (1 pL to 10 nL volumes) dispersed in an immiscible carrier oil and enables the digital manipulation of these reactors at a very high-throughput. Here, we validate a full droplet screening workflow by conducting a droplet-based cytotoxicity screen. To perform this screen, we first developed a droplet viability assay that permits the quantitative scoring of cell viability and growth within intact droplets. Next, we demonstrated the high viability of encapsulated human monocytic U937 cells over a period of 4 days. Finally, we developed an optically-coded droplet library enabling the identification of the droplets composition during the assay read-out. Using the integrated droplet technology, we screened a drug library for its cytotoxic effect against U937 cells. Taken together our droplet microfluidic platform is modular, robust, uses no moving parts, and has a wide range of potential applications including high-throughput single-cell analyses, combinatorial screening, and facilitating small sample analyses.
我们展示了一种基于液滴的微流控技术，该技术能够对单个哺乳动物细胞进行高通量筛选。这个集成平台允许将单个细胞和试剂封装在分散于不混溶的载液中的独立水性微滴（体积为1皮升至10纳升）中，并能以非常高的通量对这些反应容器进行数字操作。在此，我们通过进行基于液滴的细胞毒性筛选来验证完整的液滴筛选工作流程。为了进行此筛选，我们首先开发了一种液滴活力测定法，该方法允许对完整液滴内的细胞活力和生长进行定量评分。接下来，我们证明了封装的人单核细胞U937细胞在4天内具有高活力。最后，我们开发了一种光学编码的液滴文库，能够在测定读出过程中识别液滴组成。利用集成的液滴技术，我们筛选了一个药物文库对U937细胞的细胞毒性作用。我们的液滴微流控平台整体具有模块化且稳健的特点，无需移动部件，并且具有广泛的潜在应用，包括高通量单细胞分析、组合筛选以及促进小样本分析。
