Hu Hongxing, Eustace David, Merten Christoph A
European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg, Germany.
Lab Chip. 2015 Oct 21;15(20):3989-93. doi: 10.1039/c5lc00686d. Epub 2015 Aug 27.
The use of microfluidic droplets has become a powerful tool for the screening and manipulation of cells. However, currently this is restricted to assays involving a single cell type. Studies on the interaction of different cells (e.g. in immunology) as well as the screening of antibody-secreting cells in assays requiring an additional reporter cell, have not yet been successfully demonstrated. Based on Poisson statistics, the probability for the generation of droplets hosting exactly one cell of two different types is just 13.5%. To overcome this limitation, we have developed an approach in which different cell types are stained with different fluorescent dyes. Subsequent to encapsulation into droplets, the resulting emulsion is injected into a very compact sorting device allowing for analysis at high magnification and fixation of the cells close to the focal plane. By applying dual-color sorting, this furthermore enables the specific collection and analysis of droplets with exactly two different cells. Our approach shows an efficiency of up to 86.7% (more than 97% when also considering droplets hosting one or more cells of each type), and, hence, should pave the way for a variety of cell-based assays in droplets.
微流控液滴已成为细胞筛选和操控的有力工具。然而，目前这仅限于涉及单一细胞类型的检测。不同细胞间相互作用的研究（如在免疫学中）以及在需要额外报告细胞的检测中筛选抗体分泌细胞，尚未得到成功验证。基于泊松统计，生成恰好包含两种不同类型单个细胞的液滴的概率仅为13.5%。为克服这一限制，我们开发了一种方法，即使用不同荧光染料对不同细胞类型进行染色。在封装到液滴中后，将所得乳液注入一个非常紧凑的分选装置，该装置允许在高倍放大下进行分析，并将细胞固定在焦平面附近。通过应用双色分选，这还能够对恰好包含两种不同细胞的液滴进行特异性收集和分析。我们的方法显示出高达86.7%的效率（如果还考虑包含每种类型一个或多个细胞的液滴，效率超过97%），因此，应为各种基于液滴的细胞检测铺平道路。
