Development of a simultaneous electrorotation device with microwells for monitoring the rotation rates of multiple single cells upon chemical stimulation
Development of a simultaneous electrorotation device with microwells for monitoring the rotation rates of multiple single cells upon chemical stimulation
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开发具有微孔的同步电旋转装置，用于监测化学刺激下多个单细胞的旋转速率
DOI：
10.1039/d2lc00627h
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发表时间：
2023
期刊：
影响因子：
6.1
通讯作者：
Tomoyuki Yasukawa
中科院分区：
文献类型：
--
作者：
Masato Suzuki;Shikiho Kawai;Chean Fei Shee;Ryoga Yamada;Seiichi Uchida;Tomoyuki Yasukawa
在这里，我们描述了一种独特的同步电旋转（ROT）装置，用于通过化学刺激监测Jurkat细胞的旋转速率，而无需荧光标记和用于估计细胞旋转速率的算法。该装置包括两对交叉指状阵列电极，其通过具有矩形微孔的20 μ m厚的绝缘层正交堆叠。四个微电极（两个在微孔底部图案化，另外两个在绝缘层上）布置在矩形微孔的每一侧。当向四个微电极施加AC电压以产生旋转电场时，捕获在微孔中的细胞经历ROT。这些微孔甚至在流体流动中维持细胞。此后，在刺激期间估计并监测捕获细胞的ROT率。我们证明了通过监测细胞的ROT率来估计细胞的化学效率的可行性。在将Jurkat细胞悬浮液引入装置后，通过施加AC信号使细胞经受ROT。此外，通过添加含有离子霉素（钙离子载体）的等分试样来化学刺激旋转细胞。经离子霉素刺激后，细胞的ROT率逐渐下降，30 s后降至初始ROT率的90%。ROT率降低膜电容的增加。因此，我们的装置能够同时监测化学刺激诱导的许多细胞膜电容的变化，而无需荧光标记。
Here, we described a unique simultaneous electrorotation (ROT) device for monitoring the rotation rate of Jurkat cells via chemical stimulation without fluorescent labeling and an algorithm for estimating cell rotation rates. The device comprised two pairs of interdigitated array electrodes that were stacked orthogonally through a 20 μm-thick insulating layer with rectangular microwells. Four microelectrodes (two were patterned on the bottom of the microwells and the other two on the insulating layer) were arranged on each side of the rectangular microwells. The cells, which were trapped in the microwells, underwent ROT when AC voltages were applied to the four microelectrodes to generate a rotating electric field. These microwells maintained the cells even in fluid flows. Thereafter, the ROT rates of the trapped cells were estimated and monitored during the stimulation. We demonstrated the feasibility of estimating the chemical efficiency of cells by monitoring the ROT rates of the cells. After introducing a Jurkat cell suspension into the device, the cells were subjected to ROT by applying an AC signal. Further, the rotating cells were chemically stimulated by adding an ionomycin (a calcium ionophore)-containing aliquot. The ROT rate of the ionomycin-stimulated cells decreased gradually to 90% of the initial rate after 30 s. The ROT rate was reduced by an increase in membrane capacitance. Thus, our device enabled the simultaneous chemical stimulation-induced monitoring of the alterations in the membrane capacitances of many cells without fluorescent labeling.