A highly-occupied, single-cell trapping microarray for determination of cell membrane permeability.
A highly-occupied, single-cell trapping microarray for determination of cell membrane permeability.
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DOI：
10.1039/c7lc00883j
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发表时间：
2017-11-21
期刊：
影响因子：
6.1
通讯作者：
Toner M
中科院分区：
文献类型：
--
作者：
Weng L;Ellett F;Edd J;Wong KHK;Uygun K;Irimia D;Stott SL;Toner M
半渗透性和选择性渗透性是细胞膜的一个基本的重要特性。膜渗透性可以通过监测暴露于非等渗环境后细胞的体积变化来确定。为此，最近已经开发了几种微流体灌注室。然而，这些装置仅允许观察可能以不受控制的方式彼此相互作用的单个细胞或一组细胞。其中一些器件集成了片上温度控制，以研究膜渗透性的温度依赖性，但它们不可避免地需要复杂的制造和组装，以及精密的温度和压力校准。因此，非常需要设计一种简单的单细胞捕获装置，其允许平行监测在各种温度下经受非等渗暴露的多个单独的个体细胞。在这项研究中，我们开发了一个无泵，单层芯片与高陷阱占用的单细胞。该装置的基准性能通过靶向直径为18.8 μm的球形颗粒作为模型进行，产生高达86.8%的陷阱占有率，行到行的位移为10-30 μm。研究还表明，在每个阵列中，大于相应临界尺寸的颗粒将被陷阱以确定性的横向位移模式排除。为了证明这种方法的实用性，我们使用单细胞捕获装置在4、22和37°C下测定大鼠肝细胞和患者来源的循环肿瘤细胞（Brx-142）的膜渗透性。发现大鼠肝细胞膜通过脂质和水通道蛋白介导的途径对水和小分子如DMSO和甘油具有高度渗透性。然而，Brx-142细胞显示出比大鼠肝细胞低的膜通透性，这与水和DMSO转运的强耦合有关，但水和甘油之间的相互作用较少。本文报道的膜渗透性数据为膜转运的生物物理学提供了新的见解，如水通道蛋白表达和水与溶质的偶联转运，并为稀有细胞和珍贵组织的生物库的最终目标提供了必要的数据。一个被动泵，单细胞捕获微阵列的开发，以监测多个，单细胞的体积变化后，高渗暴露。
Semi- and selective permeability is a fundamentally important characteristic of the cell membrane. Membrane permeability can be determined by monitoring the volumetric change of cells following exposure to a non-isotonic environment. For this purpose, several microfluidic perfusion chambers have been developed recently. However, these devices only allow the observation of one single cell or a group of cells that may interact with one another in an uncontrolled way. Some of these devices have integrated on-chip temperature control to investigate the temperature-dependence of membrane permeability, but they inevitably require sophisticated fabrication and assembly, and delicate temperature and pressure calibration. Therefore, it is highly desirable to design a simple single-cell trapping device that allows parallel monitoring of multiple separate, individual cells subjected to non-isotonic exposure at various temperatures. In this study, we developed a pumpless, single-layer microarray with high trap occupancy of single cells. The benchmark performance of the device was conducted by targeting spherical particles of 18.8 μm in diameter as a model, yielding trap occupancy of up to 86.8% with a row-to-row shift of 10–30 μm. It was also revealed that in each array the particles larger than a corresponding critical size would be excluded by the traps in a deterministic lateral displacement mode. Demonstrating the utility of this approach, we used the single-cell trapping device to determine the membrane permeability of rat hepatocytes and patient-derived circulating tumor cells (Brx-142) at 4, 22 and 37°C. The membrane of rat hepatocytes was found to be highly permeable to water and small molecules such as DMSO and glycerol, via both lipid- and aquaporin-mediated pathways. Brx-142 cells, however, displayed lower membrane permeability than rat hepatocytes, which was associated with strong coupling of water and DMSO transport but less interaction between water and glycerol. The membrane permeability data reported here provide new insights into the biophysics of membrane transport such as aquaporin expression and coupling transport of water and solutes, as well as providing essential data for the ultimate goal of biobanking rare cells and precious tissues. A passive pumping, single-cell trapping microarray was developed to monitor volumetric change of multiple, single cells following hypertonic exposure.
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影响因子：
2.7
作者：
CHESNE, C;GUILLOUZO, A
通讯作者：
GUILLOUZO, A
影响因子：
2.7
作者：
Paynter, S. J.;Andrews, K. J.;Dunnett, S. B.
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Dunnett, S. B.
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56.9
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Sturm, JC
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2.9
作者：
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Marinelli, RA
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8.4
作者：
Lyu, Shaw-Ruey;Chen, Wun-Jie;Hsieh, Wen-Hsin
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Hsieh, Wen-Hsin