Hydrodynamic-based microfluidic platforms enable single-cell arraying and analysis over time. Despite the advantages of established microfluidic systems, long-term analysis and proliferation of cells selected in such devices require off-chip recovery of cells as well as an investigation of on-chip analysis on cell phenotype, requirements still largely unmet. Here, we introduce a device for single-cell isolation, selective retrieval and off-chip recovery. To this end, singularly addressable three-dimensional electrodes are embedded within a microfluidic channel, allowing the selective release of single cells from their trapping site through application of a negative dielectrophoretic (DEP) force. Selective capture and release are carried out in standard culture medium and cells can be subsequently mitigated towards a recovery well using micro-engineered hybrid SU-8/PDMS pneumatic valves. Importantly, transcriptional analysis of recovered cells revealed only marginal alteration of their molecular profile upon DEP application, underscored by minor transcriptional changes induced upon injection into the microfluidic device. Therefore, the established microfluidic system combining targeted DEP manipulation with downstream hydrodynamic coordination of single cells provides a powerful means to handle and manipulate individual cells within one device.

中文翻译：

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结合介电电泳力和定向流体动力流从微流控阵列中选择性提取单个细胞

基于水动力的微流体平台可实现随时间推移的单细胞排列和分析。尽管已建立的微流体系统具有优势，但在此类设备中选择的细胞进行长期分析和增殖仍需要芯片外恢复细胞以及对细胞表型进行芯片上分析的研究，但仍未满足要求。在这里，我们介绍一种用于单细胞隔离，选择性检索和片外恢复的设备。为此，可单寻址的三维电极嵌入微流体通道中，从而允许通过施加负介电电泳（DEP）力将单个细胞从其捕获位点选择性释放。选择性捕获和释放在标准培养基中进行，随后可以使用微工程混合SU-8 / PDMS气动阀将细胞减缓至恢复井。重要的是，对回收细胞的转录分析显示，在DEP应用后，它们的分子谱仅发生了边际变化，这一点在注入微流控设备后引起的微小转录变化中得到了强调。因此，已建立的微流体系统将靶向DEP操纵与单个细胞的下游流体动力协调相结合，提供了一种强大的手段来处理和操纵一个设备中的单个细胞。通过注射到微流体装置中引起的微小转录变化来强调。因此，已建立的微流体系统将靶向DEP操纵与单个细胞的下游流体动力协调相结合，提供了一种强大的手段来处理和操纵一个设备中的单个细胞。通过注射到微流体装置中引起的微小转录变化来强调。因此，已建立的微流体系统将靶向DEP操纵与单个细胞的下游流体动力协调相结合，提供了一种强大的手段来处理和操纵一个设备中的单个细胞。