Leonaviciene Greta, Leonavicius Karolis, Meskys Rolandas, Mazutis Linas
Institute of Biotechnology, Life Science Centre, Vilnius University, 7 Sauletekio av., Vilnius, LT-10257, Lithuania.
Institute of Biochemistry, Life Science Centre, Vilnius University, 7 Sauletekio av., Vilnius, LT-10257, Lithuania.
Lab Chip. 2020 Oct 27;20(21):4052-4062. doi: 10.1039/d0lc00660b.
Droplet microfluidics technology provides a powerful approach to isolate and process millions of single cells simultaneously. Despite many exciting applications that have emerged based on this technology, workflows based on multi-step operations, including molecular biology and cell-based phenotypic screening assays, cannot be easily adapted to droplet format. Here, we present a microfluidics-based technique to isolate single cells, or biological samples, into semi-permeable hydrogel capsules and perform multi-step biological workflows on thousands to millions of individual cells simultaneously. The biochemical reactions are performed by changing the aqueous buffer surrounding the capsules, without needing sophisticated equipment. The semi-permeable nature of the capsules' shell retains large encapsulated biomolecules (such as genome) while allowing smaller molecules (such as proteins) to passively diffuse. In contrast to conventional hydrogel bead assays, the approach presented here improves bacterial cell retention during multi-step procedures as well as the efficiency of biochemical reactions. We showcase two examples of capsule use for single genome amplification of bacteria, and expansion of individual clones into isogenic microcolonies for later screening for biodegradable plastic production.
微滴微流控技术提供了一种强大的方法，可同时分离和处理数百万个单细胞。尽管基于该技术已出现了许多令人兴奋的应用，但基于多步操作的工作流程，包括分子生物学和基于细胞的表型筛选分析，却难以轻松适应微滴形式。在此，我们展示了一种基于微流控的技术，可将单细胞或生物样本分离到半透性水凝胶胶囊中，并同时对数千至数百万个单个细胞执行多步生物工作流程。通过改变胶囊周围的水性缓冲液来进行生化反应，无需复杂的设备。胶囊外壳的半透性可保留被封装的大分子（如基因组），同时允许小分子（如蛋白质）被动扩散。与传统的水凝胶珠分析相比，本文介绍的方法提高了多步程序中细菌细胞的保留率以及生化反应的效率。我们展示了两个使用胶囊进行细菌单基因组扩增以及将单个克隆扩展为同基因微菌落以便后续筛选可生物降解塑料生产的示例。
