Department of Chemistry, Iowa State University, 2415 Osborn Dr., 1605 Gilman Hall, Ames, IA, 50011, USA.
Anal Bioanal Chem. 2018 Apr;410(10):2499-2515. doi: 10.1007/s00216-018-0896-y. Epub 2018 Feb 23.
In this review, recent advances that leverage dielectrophoretic approaches to accomplish single-cell analysis (both "on-chip" and "off-chip") are discussed with special emphasis on eukaryotic cells. Dielectrophoresis as an electric-field-induced force utilized for cell manipulation can confer selectivity without labeling. Recent technical improvements have increased the volumetric throughput of the separation of cells from complex mixtures, introduced new strategies for massively parallel single-cell confinement for subsequent on-chip analysis, made possible selective transport of individual cells off-chip, and integrated preconcentration and prefocusing steps to enhance dielectrophoretic performance. Collectively, these studies potentiate all-in-one platforms capable of taking as their input complex mixtures of cells and accomplishing single-cell analysis. Assays requiring small reaction volumes (e.g., enzymatic assays, fluorescent in situ hybridization, and immunostaining) have been demonstrated. Still greater opportunities to unravel cell-to-cell variations and for point-of-care applications can be realized by making possible on-chip gene amplification, live-cell assays, and either dielectrophoretic manipulation in native media or on-chip exchange of media. We therefore conclude with a discussion of emerging capabilities in these areas.
在这篇综述中,我们讨论了利用介电泳方法进行单细胞分析(包括“片上”和“片外”)的最新进展,特别强调了真核细胞。介电泳作为一种用于细胞操作的电场诱导力,可以在不进行标记的情况下赋予选择性。最近的技术改进提高了从复杂混合物中分离细胞的体积通量,引入了用于大规模并行单细胞限制的新策略,以进行后续的片上分析,实现了单个细胞的选择性片外输送,并集成了预浓缩和预聚焦步骤,以增强介电泳性能。总的来说,这些研究为能够将复杂的细胞混合物作为输入并完成单细胞分析的一体化平台提供了可能性。已经证明了需要小反应体积的测定(例如,酶测定、荧光原位杂交和免疫染色)。通过实现片上基因扩增、活细胞测定以及在天然介质中或片上进行介电泳操作或介质交换,可以实现更大的机会来揭示细胞间的变化,并实现即时护理应用。因此,我们最后讨论了这些领域的新兴能力。