A high-performance elastomeric patch clamp chip
A high-performance elastomeric patch clamp chip
复制标题
DOI：
10.1039/b607913j
复制
发表时间：
2006-10-01
期刊：
影响因子：
6.1
通讯作者：
Folch, Albert
中科院分区：
文献类型：
--
作者：
Chen, Chihchen;Folch, Albert
离子通道在细胞生理学中起着关键作用，并且是多种疾病的基础。直到今天，研究离子通道的黄金标准是膜片钳技术。膜片钳包括将细尖玻璃微量移液器小心定位在细胞表面上以形成高电阻（> 1G Ω）密封（“gigaseal”），这是一种费力、振动敏感且不易于自动化的过程。此外，移液器内的溶液不易更换。最近报道的膜片钳芯片提供了增加吞吐量的潜力，但迄今为止，与移液管相比，大多数设计的整体每细胞性能非常低，和/或制造工艺过于昂贵。在这里，我们展示了一个复制成型的弹性膜片钳芯片纳入纳米制造的收缩，提供高稳定性gigaseals，成功率与移液管，使用大鼠嗜碱性白血病（RBL）细胞。高稳定性使细胞外和细胞内的溶液在全细胞记录过程中交换。在103个实验的样本中，66个细胞（64%）被成功地固定在贴片孔处; 38个细胞（58%的固定化细胞，所有细胞的37%）被成功地gigasealed;并且25个细胞（65%的gigasealed细胞，34%的固定化细胞，所有细胞的24%）被成功地穿孔用于全细胞获取。在最后一组的27个实验中，79%的细胞可以被固定，其中68%可以被gigasealed和46%穿孔用于全细胞访问，这表明灵活性很重要。
Ion channels play key roles in cell physiology and underlie a broad spectrum of disorders. To this day, the gold standard for studying ion channels is the patch clamp technique. Patch clamping involves careful positioning of a fine-tipped glass micropipette onto the surface of the cell to form a high-resistance (> I G Omega) seal ("gigaseal"), a procedure that is laborious, vibration-sensitive, and not easily amenable to automation. In addition, the solution inside the pipette cannot be easily exchanged. Recently reported patch clamp chips offer the potential of increased throughput, but to date the overall per-cell performance of most designs has been very low when compared to pipettes, and/or the fabrication process is prohibitively expensive. Here we demonstrate a replica-molded elastomeric patch clamp chip incorporating nanofabricated constrictions, which delivers high-stability gigaseals, with success rates comparable to those of pipettes, using rat basophilic leukemia (RBL) cells. The high stability enables exchanges of both the extracellular and intracellular solution during whole-cell recordings. In a sample of 103 experiments, 66 cells (64%) were successfully immobilized at the patch aperture; 38 cells (58% of immobilized cells, 37% of all cells) were successfully gigasealed; and 25 cells (65% of gigasealed cells, 34% of immobilized cells, 24% of all cells) were successfully perforated for whole-cell access. In the last group of 27 experiments, 79% of the cells could be immobilized, of which 68% could be gigasealed and 46% perforated for whole-cell access, indicating that dexterity is important.