Separation of Cells and Microparticles in Insulator-Based Electrokinetic Systems
Separation of Cells and Microparticles in Insulator-Based Electrokinetic Systems
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基于绝缘体的动电系统中细胞和微粒的分离
DOI：
10.1021/acs.analchem.2c04366
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发表时间：
2023
影响因子：
7.4
通讯作者：
Lapizco-Encinas, Blanca H.
中科院分区：
文献类型：
--
作者：
Vaghef-Koodehi, Alaleh;Ernst, Olivia D.;Lapizco-Encinas, Blanca H.
本文介绍了在基于绝缘体的电动力（iEK）系统中，利用线性和非线性电动力现象首次连续分离具有相似特征的微粒和细胞。通过利用具有绝缘特性的设备，使电场分布扭曲，可以将线性和非线性EK现象结合起来，从而产生利用非线性电泳新进展的高效分离方案。这项工作结合了数学建模和实验来分离四种不同的粒子和细胞的二元混合物。利用COMSOL Multiphysics计算模型预测了iEK器件中粒子和细胞的滞留时间（tR,p）。然后，在模型确定的条件下进行实验分离，测量颗粒和细胞的实验保留时间（tR,e）。通过增加难度，共进行了四种不同的二元混合物分离。在第一次分离中，分离了两种类型的聚苯乙烯微颗粒，分别用于模拟大肠杆菌和酿酒酵母细胞。通过利用从第一次分离中收集到的知识，不同域和显著大小差异的细胞的混合物，E。coliandS。酿酒酵母，成功分离。第三种分离也有不同结构域的细胞，但大小更接近：蜡样芽孢杆菌与芽孢杆菌。酵母。最后一次分离包括同一结构域和属的细胞，B。cereusversusBacillus细小。以板数(N)和分离分辨率（Rs）对分离结果进行评价，其中所有分离的Rs值均大于1.5，说明分离完全。实验结果与模型结果在保留时间上一致，偏差在6-27%之间，而重复次数的差异在2 - 18%之间，具有良好的再现性。本报告首次预测了细胞在iEK系统中的滞留时间。
Presented here is the first continuous separation of microparticles and cells of similar characteristics employing linear and nonlinear electrokinetic phenomena in an insulator-based electrokinetic (iEK) system. By utilizing devices with insulating features, which distort the electric field distribution, it is possible to combine linear and nonlinear EK phenomena, resulting in highly effective separation schemes that leverage the new advancements in nonlinear electrophoresis. This work combines mathematical modeling and experimentation to separate four distinct binary mixtures of particles and cells. A computational model with COMSOL Multiphysics was used to predict the retention times (tR,p) of the particles and cells in iEK devices. Then, the experimental separations were carried out using the conditions identified with the model, where the experimental retention time (tR,e) of the particles and cells was measured. A total of four distinct separations of binary mixtures were performed by increasing the level of difficulty. For the first separation, two types of polystyrene microparticles, selected to mimicEscherichia coliandSaccharomyces cerevisiaecells, were separated. By leveraging the knowledge gathered from the first separation, a mixture of cells of distinct domains and significant size differences,E. coliandS. cerevisiae, was successfully separated. The third separation also featured cells of different domains but closer in size:Bacillus cereusversusS. cerevisiae. The last separation included cells in the same domain and genus,B. cereusversusBacillus subtilis. Separation results were evaluated in terms of number of plates (N) and separation resolution (Rs), whereRsvalues for all separations were above 1.5, illustrating complete separations. Experimental results were in agreement with modeling results in terms of retention times, with deviations in the 6–27% range, while the variation between repetitions was between 2 and 18%, demonstrating good reproducibility. This report is the first prediction of the retention time of cells in iEK systems.