Department of Mechanical and Aerospace Engineering, SUNY-Buffalo, Buffalo, New York 14260, USA.
Biomicrofluidics. 2012 Sep 24;6(3):34121. doi: 10.1063/1.4754599. eCollection 2012.
In this paper, we present a microfluidic chip that is capable of measuring electrical conductance through gap junction channels in a 2-dimensional cell sheet. The chip utilizes a tri-stream laminar flow to create a non-conductive sucrose gap between the two conducting solutions so that electrical current can pass across the sucrose gap only through the cells. Using the chip, we tested the effect of a gap junction inhibitor, 2-APB, on the electrical coupling of connexin 43 (Cx43) gap junction channels in NRK-49F cells. We found that 2-APB reversibly blocks the conductivity in a dose-dependent manner. The tri-stream chip further allows us to simultaneously follow the conductance changes and dye diffusion in real time. We show that 2-APB affects both conductance and diffusion, supporting the interpretation that both sets of data reflect the same gap junction activity. The chip provides a generic platform to investigate gap junction properties and to screen drugs that may inhibit or potentiate gap junction transmission.
在本文中,我们提出了一种微流控芯片,能够测量二维细胞片层中缝隙连接通道的电导。该芯片利用三股层流在两个导电溶液之间产生非导电蔗糖间隙,使得电流只能通过细胞穿过蔗糖间隙。使用该芯片,我们测试了缝隙连接抑制剂 2-APB 对 NRK-49F 细胞中连接蛋白 43 (Cx43) 缝隙连接通道电耦联的影响。我们发现 2-APB 以剂量依赖的方式可逆地阻断电导率。三股流芯片还允许我们实时同时跟踪电导变化和染料扩散。我们表明 2-APB 同时影响电导和扩散,支持这两组数据均反映相同的缝隙连接活动的解释。该芯片为研究缝隙连接特性和筛选可能抑制或增强缝隙连接传递的药物提供了通用平台。