A Microfluidic Pipette for Single-Cell Pharmacology
A Microfluidic Pipette for Single-Cell Pharmacology
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DOI：
10.1021/ac100480f
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发表时间：
2010-06-01
影响因子：
7.4
通讯作者：
Jesorka, Aldo
中科院分区：
文献类型：
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作者：
Ainla, Alar;Jansson, Erik T.;Jesorka, Aldo
我们报告了一种由聚二甲基硅氧烷制成的独立式微流体移液器，其循环液体尖端在出口通道前产生自约束体积。该方法灵活且可扩展，因为再循环区域的几何形状和大小由压力、通道数量和几何形状定义。移液管能够在细胞和组织培养物中的选定细胞上执行各种复杂的流体处理操作，例如混合、多路复用或梯度生成。通过摄取实验，我们发现可以从表达质子激活的人瞬时受体电位香草样蛋白（hTRPV1）受体的贴壁中国仓鼠卵巢细胞原位生成剂量反应曲线。利用密闭的细胞灌注和细胞刺激，我们可以激活单个细胞中的hTRPV1受体，通过膜片钳移液管测量反应，并通过将选定的细胞组分别暴露于甲醛/含二硫苏糖醇的溶液中诱导膜泡形成。简而言之，微流控移液管允许复杂，无污染的多种化合物输送，用于完整贴壁细胞的药理学筛选。
We report on a free-standing microfluidic pipette made in poly(dimethylsiloxane) having a circulating liquid tip that generates a self-con-fining volume in front of the outlet channels. The method is flexible and scalable as the geometry and the size of the recirculation zone is defined by pressure, channel number, and geometry. The pipette is capable of carrying out a variety of complex fluid processing operations, such as mixing, multiplexing, or gradient generation at selected cells in cell and tissue cultures. Using an uptake assay, we show that it is possible to generate dose response curves in situ from adherent Chinese hamster ovary cells expressing proton-activated human transient receptor potential vanilloid (hTRPV1) receptors. Using confined superfusion and cell stimulation, we could activate hTRPV1 receptors in single cells, measure the response by a patch-clamp pipette, and induce membrane bleb formation by exposing selected groups of cells to formaldehyde/dithiothreitol-containing solutions, respectively. In short, the microfluidic pipette allows for complex, contamination-free multiple-compound delivery for pharmacological screening of intact adherent cells.