Gorelik Julia, Gu Yuchun, Spohr Hilmar A, Shevchuk Andrew I, Lab Max J, Harding Sian E, Edwards Christopher R W, Whitaker Michael, Moss Guy W J, Benton David C H, Sánchez Daniel, Darszon Alberto, Vodyanoy Igor, Klenerman David, Korchev Yuri E
Division of Medicine, Imperial College of Science, Technology and Medicine, MRC Clinical Sciences Centre, DuCane Road, London W12 0NN, United Kingdom.
Biophys J. 2002 Dec;83(6):3296-303. doi: 10.1016/S0006-3495(02)75330-7.
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.
我们开发了一种扫描膜片钳技术，该技术有助于从传统方法无法触及的小细胞和亚微米级细胞结构进行单通道记录。扫描膜片钳技术通过单个玻璃纳米吸管探针将扫描离子电导显微镜和膜片钳记录相结合。在这种方法中，首先使用电流反馈在细胞表面扫描纳米吸管，以获得高分辨率的地形图。然后使用同一吸管进行膜片钳记录。由于图像信息是通过膜片电极获得的，因此可用于将吸管以纳米精度定位到细胞上。通过从上皮微绒毛顶部和心肌细胞T小管开口处获得离子通道记录，证明了该技术的实用性。此外，我们首次证明，在生理条件下从非常小的细胞（如精子细胞）记录离子通道以及从亚微米级神经元突起等细胞微观结构记录离子通道是可能的。
