Dielectrophoresis-Assisted Self-Digitization Chip for High-Efficiency Single-Cell Analysis.
Dielectrophoresis-Assisted Self-Digitization Chip for High-Efficiency Single-Cell Analysis.
复制标题
用于高效单细胞分析的介电泳辅助自数字化芯片。
DOI：
10.1007/978-1-0716-3323-6_3
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Chiu,DanielT
中科院分区：
文献类型：
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作者：
Qin,Yuling;Wu,Li;Chiu,DanielT
单细胞分析细胞表型信息，如表面蛋白表达和核酸含量，对于了解细胞群体内的异质性是必不可少的。这里描述了一种介电辅助自数字化(SD)微流控芯片的设计和使用；它在隔离的微室中高效地捕获单细胞以进行单细胞分析。这种自数字化芯片通过流体力、界面张力和通道几何形状的组合，自发地将水溶液分割成微腔。由于外加交流电压产生的局部电场极大值，单个细胞通过介电(DEP)被引导并捕获在微室的入口处。多余的细胞被冲走，捕获的细胞被释放到腔室中，并通过关闭外部电压、通过芯片运行反应缓冲液以及通过周围通道的不相容油相流动密封腔室来准备进行原位分析。通过基于环介导的等温扩增(LAMP)的单细胞核酸定量，展示了该装置在单细胞分析中的应用。该平台为药物发现相关的单细胞研究提供了一个强大的新工具。例如，从数字芯片观察到的癌症相关突变基因的单细胞基因分型可能是靶向治疗的有用生物标记物。
Single-cell analysis of cell phenotypic information such as surface protein expression and nucleic acid content is essential for understanding heterogeneity within cell populations. Here the design and use of a dielectrophoresis-assisted self-digitization (SD) microfluidics chip is described; it captures single cells in isolated microchambers with high efficiency for single-cell analysis. The self-digitization chip spontaneously partitions aqueous solution into microchambers through a combination of fluidic forces, interfacial tension, and channel geometry. Single cells are guided to and trapped at the entrances of microchambers by dielectrophoresis (DEP) due to local electric field maxima created by an externally applied AC voltage. Excess cells are flushed away, and trapped cells are released into the chambers and prepared for in situ analysis by turning off the external voltage, by running reaction buffer through the chip, and by sealing the chambers with a flow of an immiscible oil phase through the surrounding channels. The use of this device in single-cell analysis is demonstrated by performing single-cell nucleic acid quantitation based on loop-mediated isothermal amplification (LAMP). This platform provides a powerful new tool for single-cell research pertaining to drug discovery. For example, the single-cell genotyping of cancer-related mutant gene observed from the digital chip could be useful biomarker for targeted therapy.