Xu Yeye, Ding Weiping, Li Shibo, Li Chengpan, Gao Dayong, Qiu Bensheng
Center for Biomedical Engineering, University of Science and Technology of China, Hefei, 230027, Anhui, China.
Department of Electronic Science and Technology, University of Science and Technology of China, Hefei, 230027, Anhui, China.
Anal Bioanal Chem. 2020 Jul;412(19):4537-4548. doi: 10.1007/s00216-020-02704-7. Epub 2020 May 21.
The hydraulic permeability of the lipid bilayer membrane of a single cell, a very important parameter in biological and medical fields, has been attracting increasing attention. To date, methods developed to determine this permeability are either operation-complicated or time-consuming. Therefore, we developed a chip for automatically and rapidly determining the permeability of cells that integrates microfluidics and cell impedance analysis. The chip is designed to automatically identify a single cell, capture the cell, and record the volume change in that cell. We confirmed the abilities of single-cell identification and capture with the upper and lower voltage thresholds determined, validated the performance of the differential electrode design for accurate cell volume measurements, deduced the extracellular osmotic pressure change in the presence of a hypertonic solution according to fluorescence intensity, and demonstrated the single-cell volume change recorded by the chip. Then, the accuracy of the permeability determined with the chip was verified using HeLa cells. Finally, the permeability of human-induced pluripotent stem cells (hiPSCs) was determined to be 0.47 ± 0.03 μm/atm/min. Using the chip, the permeability can be determined within 5 min. This study provides insights for the new design of an automatic single-cell identification and capture chip for single cell-related studies. Graphical abstract.
单细胞脂质双分子层膜的水力渗透性是生物和医学领域中一个非常重要的参数，一直备受关注。迄今为止，用于测定这种渗透性的方法要么操作复杂，要么耗时。因此，我们开发了一种集成微流控和细胞阻抗分析的芯片，用于自动快速测定细胞的渗透性。该芯片旨在自动识别单个细胞、捕获细胞并记录该细胞的体积变化。我们通过确定的上下电压阈值确认了单细胞识别和捕获能力，验证了用于精确细胞体积测量的差分电极设计的性能，根据荧光强度推导了高渗溶液存在时的细胞外渗透压变化，并展示了芯片记录的单细胞体积变化。然后，使用HeLa细胞验证了用该芯片测定的渗透性的准确性。最后，测定人诱导多能干细胞（hiPSC）的渗透性为0.47±0.03μm/atm/min。使用该芯片，可在5分钟内测定渗透性。本研究为用于单细胞相关研究的自动单细胞识别和捕获芯片的新设计提供了思路。图形摘要。
