Ion channels in small cells and subcellular structures can be studied with a smart patch-clamp system
Ion channels in small cells and subcellular structures can be studied with a smart patch-clamp system
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DOI：
10.1016/s0006-3495(02)75330-7
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发表时间：
2002-12-01
影响因子：
3.4
通讯作者：
Korchev, YE
中科院分区：
文献类型：
--
作者：
Gorelik, J;Gu, YC;Korchev, YE
我们已经开发了一种扫描膜片钳技术，可以方便地从传统方法无法进入的小细胞和亚微米细胞结构中进行单通道记录。扫描膜片钳技术结合了扫描离子电导显微镜和通过单个玻璃纳米探针进行膜片钳记录的技术。在这种方法中，纳米吸管首先在细胞表面扫描，使用电流反馈，以获得高分辨率的地形图像。然后使用相同的移液器进行膜片钳记录。由于图像信息是通过贴片电极获得的，因此它可以用于以纳米精度将移液器定位到细胞上。通过从上皮微绒毛顶部和心肌细胞t小管开口获得离子通道记录，证明了该技术的实用性。此外，我们首次证明了在生理条件下记录非常小的细胞（如精子细胞）的离子通道以及记录细胞微观结构（如亚微米神经元过程）是可能的。
We have developed a scanning patch-clamp technique that facilitates single-channel recording from small cells and submicron cellular structures that are inaccessible by conventional methods. The scanning patch-clamp technique combines scanning ion conductance microscopy and patch-clamp recording through a single glass nanopipette probe. In this method the nanopipette is first scanned over a cell surface, using current feedback, to obtain a high-resolution topographic image. This same pipette is then used to make the patch-clamp recording. Because image information is obtained via the patch electrode it can be used to position the pipette onto a cell with nanometer precision. The utility of this technique is demonstrated by obtaining ion channel recordings from the top of epithelial microvilli and openings of cardiomyocyte T-tubules. Furthermore, for the first time we have demonstrated that it is possible to record ion channels from very small cells, such as sperm cells, under physiological conditions as well as record from cellular microstructures such as submicron neuronal processes.