微滴微流控技术彻底改变了生物分子分析研究,因为它有能力保留基因型与表型的联系,并有助于揭示异质性。大量且均匀的皮升液滴的特征是将溶液分裂到每个液滴中的单细胞和单分子可以可视化的水平,条形码,并分析。然后,液滴分析可以展开密集的基因组数据,提供高灵敏度,并从大量的组合或表型中进行筛选和排序。基于这些独特的优势,这篇综述的重点是关于利用微滴微流控技术的各种筛选应用的最新研究。首先介绍了液滴微流控技术的新兴进展,包括高效和扩大液滴封装,和普遍的批量操作。然后简要研究了基于液滴的数字检测测定和单细胞突变测序的新实现,以及相关应用,如药物敏感性测试,用于癌症亚型鉴定的多路复用,病毒与宿主的相互作用,以及多模态和时空分析。同时,我们专注于基于液滴的大规模组合筛选所需的表型,强调免疫细胞的分选,抗体,酶学性质,和通过定向进化方法产生的蛋白质。最后,一些挑战,还讨论了液滴微流体技术在实践中的部署和未来前景。
Droplet microfluidic technology has revolutionized biomolecular analytical research, as it has the capability to reserve the genotype-to-phenotype linkage and assist for revealing the heterogeneity. Massive and uniform picolitre droplets feature dividing solution to the level that single cell and single molecule in each droplet can be visualized, barcoded, and analyzed. Then, the droplet assays can unfold intensive genomic data, offer high sensitivity, and screen and sort from a large number of combinations or phenotypes. Based on these unique advantages, this review focuses on up-to-date research concerning diverse screening applications utilizing droplet microfluidic technology. The emerging progress of droplet microfluidic technology is first introduced, including efficient and scaling-up in droplets encapsulation, and prevalent batch operations. Then the new implementations of droplet-based digital detection assays and single-cell muti-omics sequencing are briefly examined, along with related applications such as drug susceptibility testing, multiplexing for cancer subtype identification, interactions of virus-to-host, and multimodal and spatiotemporal analysis. Meanwhile, we specialize in droplet-based large-scale combinational screening regarding desired phenotypes, with an emphasis on sorting for immune cells, antibodies, enzymatic properties, and proteins produced by directed evolution methods. Finally, some challenges, deployment and future perspective of droplet microfluidics technology in practice are also discussed.