Radboud University, Institute for Molecules and Materials, Heyendaalseweg 135, 6525AJ, Nijmegen, The Netherlands.
Adv Biosyst. 2020 Jan;4(1):e1900188. doi: 10.1002/adbi.201900188. Epub 2019 Nov 26.
Droplet microfluidics has revolutionized the study of single cells. The ability to compartmentalize cells within picoliter droplets in microfluidic devices has opened up a wide range of strategies to extract information at the genomic, transcriptomic, proteomic, or metabolomic level from large numbers of individual cells. Studying the different molecular landscapes at single-cell resolution has provided the authors with a detailed picture of intracellular heterogeneity and the resulting changes in cellular phenotypes. In addition, these technologies have aided in the discovery of rare cells in tumors or in the immune system, and left the authors with a deeper understanding of the fundamental biological processes that determine cell fate. This review aims to provide a detailed overview of the various droplet microfluidic strategies reported in the literature, taking into account the sometimes subtle differences in workflow or reagents that enable or improve certain protocols. Specifically, approaches to targeted- and whole-genome analysis, as well as whole-transcriptome profiling techniques, are reviewed. In addition, an up-to-date overview of new methods to characterize and quantify single-cell protein levels, and of developments to screen secreted molecules such as antibodies, cytokines, or metabolites at the single-cell level, is provided.
液滴微流控技术已经彻底改变了单细胞研究。在微流控设备中将细胞分隔在皮升级别的液滴中的能力，为从大量单个细胞中提取基因组、转录组、蛋白质组或代谢组水平的信息提供了广泛的策略。研究单细胞分辨率下的不同分子图谱，使作者能够详细了解细胞内异质性以及由此导致的细胞表型变化。此外，这些技术有助于发现肿瘤或免疫系统中的稀有细胞，并使作者更深入地了解决定细胞命运的基本生物学过程。本文综述旨在详细概述文献中报道的各种液滴微流控策略，同时考虑到有时在工作流程或试剂方面存在细微差异，这些差异可以实现或改进某些方案。具体来说，本文综述了靶向和全基因组分析方法，以及全转录组分析技术。此外，本文还提供了单细胞蛋白水平的特征和定量的新方法的最新概述，以及在单细胞水平上筛选抗体、细胞因子或代谢物等分泌分子的新进展。
