Chemical Transfection of Cells in Picoliter Aqueous Droplets in Fluorocarbon Oil
Chemical Transfection of Cells in Picoliter Aqueous Droplets in Fluorocarbon Oil
复制标题
氟碳油中皮升水滴中细胞的化学转染
DOI：
10.1021/ac2022794
复制
发表时间：
2011-11-15
影响因子：
7.4
通讯作者：
Lu, Chang
中科院分区：
文献类型：
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作者：
Chen, Fangyuan;Zhan, Yihong;Lu, Chang
油包水液滴内的细胞操作对于使用液滴微流体进行基于细胞的测定的高通量筛选至关重要。液滴内的细胞转染是功能基因组学研究的关键步骤，该研究使用液滴平台检查基因的原位功能。传统的烃包水油滴与化学转染不相容，因为它会损害细胞活力并从水相中萃取有机转染试剂。在这项工作中，我们研究了封装在氟碳油皮升液滴中的细胞的化学转染。使用氟碳油可实现高细胞活力，并且转染试剂进入油相的损失很少。我们改变了液滴内的孵育时间、DNA 浓度和液滴大小。经过优化后，我们能够在液滴中实现与本体溶液中相似的转染效率。有趣的是，转染效率随液滴变小而增加，这表明微尺度限制或表面积与体积比的影响。
The manipulation of cells inside water-in-oil droplets is essential for high-throughput screening of cell-based assays using droplet microfluidics. Cell transfection inside droplets is a critical step involved in functional genomics studies that examine in situ functions of genes using the droplet platform. Conventional water-in-hydrocarbon oil droplets are not compatible with chemical transfection due to its damage to cell viability and extraction of organic transfection reagents from the aqueous phase. In this work, we studied chemical transfection of cells encapsulated in picoliter droplets in fluorocarbon oil. The use of fluorocarbon oil permitted high cell viability and little loss of the transfection reagent into the oil phase. We varied the incubation time inside droplets, the DNA concentration, and the droplet size. After optimization, we were able to achieve similar transfection efficiency in droplets to that in the bulk solution. Interestingly, the transfection efficiency increased with smaller droplets, suggesting effects from either the microscale confinement or the surface-to-volume ratio.