Dynamic monitoring of single cell lysis in an impedance-based microfluidic device.
Dynamic monitoring of single cell lysis in an impedance-based microfluidic device.
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DOI：
10.1007/s10544-016-0081-z
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发表时间：
2016-08
影响因子：
2.8
通讯作者：
Seshia AA
中科院分区：
文献类型：
--
作者：
Zhou Y;Basu S;Laue ED;Seshia AA
证明了能够捕获和感测单个细胞的电特性的动态变化的微流体装置。该设备被应用于实时记录的阻抗测量的小鼠胚胎干细胞（mESC）在膜裂解的过程中，与所产生的变化，在这个过程中的细胞的电特性被定量跟踪随着时间的推移。据观察，细胞膜裂解后的阻抗幅度急剧下降。在这个过程的时间过程中，也观察到一个显着的相移谱。通过将实验数据拟合到物理模型，可以提取电池的电参数并量化过程中的参数变化。在细胞裂解实验中，发现细胞膜的等效电导率由于裂解期间膜中的孔形成而显著增加。还观察到膜的比电容的增加。另一方面，观察到细胞质的电导率降低，这可以解释为在裂解期间过量的水通过膜的逐渐透化进入细胞的事实。细胞可以被捕获在设备中长达数天，并且可以通过无标记和非侵入性方式的实时阻抗测量来监测它们的电响应。此外，由于该装置的高效单细胞捕获能力，许多细胞可以被捕获并保持在单独的威尔斯孔中用于并行平行实验，从而允许阶梯式参数实验的可能性，并通过组合整个阵列的测量来研究细胞异质性。
A microfluidic device that is capable of trapping and sensing dynamic variations in the electrical properties of individual cells is demonstrated. The device is applied to the real-time recording of impedance measurements of mouse embryonic stem cells (mESCs) during the process of membrane lysis, with the resulting changes in the electrical properties of cells during this process being quantitatively tracked over time. It is observed that the impedance magnitude decreases dramatically after cell membrane lysis. A significant shift in the phase spectrum is also observed during the time course of this process. By fitting experimental data to physical models, the electrical parameters of cells can be extracted and parameter variations quantified during the process. In the cell lysis experiments, the equivalent conductivity of the cell membrane is found to increase significantly due to pore formation in the membrane during lysis. An increase in the specific capacitance of the membrane is also observed. On the other hand, the conductivity of the cytoplasm is observed to decrease, which may be explained the fact that excess water enters the cell through the gradual permeabilization of the membrane during lysis. Cells can be trapped in the device for periods up to several days, and their electrical response can be monitored by real-time impedance measurements in a label-free and non-invasive manner. Furthermore, due to the highly efficient single cell trapping capacity of the device, a number of cells can be trapped and held in separate wells for concurrent parallel experiments, allowing for the possibility of stepped parametric experiments and studying cell heterogeneity by combining measurements across the array.