Clausell-Tormos Jenifer, Lieber Diana, Baret Jean-Christophe, El-Harrak Abdeslam, Miller Oliver J, Frenz Lucas, Blouwolff Joshua, Humphry Katherine J, Köster Sarah, Duan Honey, Holtze Christian, Weitz David A, Griffiths Andrew D, Merten Christoph A
Institut de Science et d'Ingénierie Supramoléculaires, Université Louis Pasteur, Strasbourg Cedex, France.
Chem Biol. 2008 May;15(5):427-37. doi: 10.1016/j.chembiol.2008.04.004.
High-throughput, cell-based assays require small sample volumes to reduce assay costs and to allow for rapid sample manipulation. However, further miniaturization of conventional microtiter plate technology is problematic due to evaporation and capillary action. To overcome these limitations, we describe droplet-based microfluidic platforms in which cells are grown in aqueous microcompartments separated by an inert perfluorocarbon carrier oil. Synthesis of biocompatible surfactants and identification of gas-permeable storage systems allowed human cells, and even a multicellular organism (C. elegans), to survive and proliferate within the microcompartments for several days. Microcompartments containing single cells could be reinjected into a microfluidic device after incubation to measure expression of a reporter gene. This should open the way for high-throughput, cell-based screening that can use >1000-fold smaller assay volumes and has approximately 500x higher throughput than conventional microtiter plate assays.
高通量细胞分析需要小样本体积,以降低分析成本并实现快速的样本处理。然而,由于蒸发和毛细作用,传统微孔板技术的进一步小型化存在问题。为克服这些限制,我们描述了基于液滴的微流控平台,其中细胞在由惰性全氟碳载体油分隔的水性微隔室中生长。生物相容性表面活性剂的合成和透气储存系统的鉴定使得人类细胞甚至多细胞生物(秀丽隐杆线虫)能够在微隔室中存活并增殖数天。孵育后,含有单细胞的微隔室可重新注入微流控装置中以测量报告基因的表达。这应为高通量细胞筛选开辟道路,该筛选可使用比传统微孔板分析小1000倍以上的分析体积,且通量约高500倍。