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在膜片钳实验中进行全细胞记录的一种新方法。

A novel way to go whole-cell in patch-clamp experiments.

作者信息

Inayat Samsoon, Zhao Yan, Cantrell Donal R, Dikin Dmitryi, Pinto Lawrence H, Troy John B

出版信息

IEEE Trans Biomed Eng. 2010 Nov;57(11). doi: 10.1109/TBME.2010.2055055. Epub 2010 Jun 28.

Abstract

With a conventional patch-clamp electrode, an Ag/AgCl wire sits stationary inside the pipette. To move from the gigaseal cell-attached configuration to whole-cell recording, suction is applied inside the pipette. We have designed and developed a novel Pushpen patch-clamp electrode, in which a W wire insulated and wound with Ag/AgCl wire can move linearly inside the pipette. The W wire has a conical tip, which can protrude from the pipette tip like a push pen, a procedure we call the Pushpen Operation. We use the Pushpen operation to impale the cell membrane in cell-attached configuration to go whole-cell without disruption of the gigaseal. We successfully recorded whole-cell currents from chinese hamster ovarian cells expressing influenza A virus protein A/M2, after obtaining whole-cell configuration with the Pushpen operation. This novel method of achieving whole-cell configuration may have a higher success rate than is the case with the conventional patch clamp technique.

摘要

使用传统的膜片钳电极时,一根Ag/AgCl线固定在移液管内部。要从千兆封接细胞贴附配置转换为全细胞记录,需在移液管内部施加吸力。我们设计并开发了一种新型的推笔式膜片钳电极,其中一根用Ag/AgCl线绝缘并缠绕的钨丝可以在移液管内线性移动。钨丝有一个锥形尖端,它可以像推笔一样从移液管尖端伸出,我们将这个过程称为推笔操作。我们使用推笔操作在细胞贴附配置下刺穿细胞膜以进入全细胞状态,而不会破坏千兆封接。在用推笔操作获得全细胞配置后,我们成功记录了表达甲型流感病毒蛋白A/M2的中国仓鼠卵巢细胞的全细胞电流。这种实现全细胞配置的新方法可能比传统膜片钳技术具有更高的成功率。

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