Whole cell patch clamp recording performed on a planar glass chip
Whole cell patch clamp recording performed on a planar glass chip
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DOI：
10.1016/s0006-3495(02)75646-4
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发表时间：
2002-06-01
影响因子：
3.4
通讯作者：
Behrends, JC
中科院分区：
文献类型：
--
作者：
Fertig, N;Blick, RH;Behrends, JC
膜片钳技术是研究离子通道的最新技术。离子通道介导电流流动，在细胞生理中起着至关重要的作用，是重要的药物靶点。该技术最流行的（全细胞）变体检测整个细胞膜上的集合电流。贴片夹紧仍然是一个费力的过程，需要一个熟练的实验人员在显微镜下微操作玻璃移液器，一次记录一个细胞。在这里，我们报告了一种平面，微结构石英芯片，用于全细胞膜片钳测量，无需微操作或视觉控制。用湿法蚀刻技术在厚度为200am的石英衬底上穿孔，得到直径约为1um的孔径。小孔取代了通常用于膜片钳记录的玻璃移液管的尖端。通过吸力将细胞定位到悬浮孔上。来自不同细胞类型（CHO， N1E-115神经母细胞瘤）的全细胞记录使用研究K+通道和电压门控Ca2+通道的微结构芯片进行。
The state of the art technology for the study of ion channels is the patch clamp technique. Ion channels mediate electrical current flow, have crucial roles in cellular physiology, and are important drug targets. The most popular (whole cell) variant of the technique detects the ensemble current over the entire cell membrane. Patch clamping is still a laborious process, requiring a skilled experimenter to micromanipulate a glass pipette under a microscope to record from one cell at a time. Here we report on a planar, microstructured quartz chip for whole cell patch clamp measurements without micromanipulation or visual control. A quartz substrate of 200 Am thickness is perforated by wet etching techniques resulting in apertures with diameters of similar to1 mum. The apertures replace the tip of glass pipettes commonly used for patch clamp recording. Cells are positioned onto the apertures from suspension by application of suction. Whole cell recordings from different cell types (CHO, N1E-115 neuroblastoma) are performed with microstructured chips studying K+ channels and voltage gated Ca2+ channels.