The use of scanning ion conductance microscopy to image A6 cells
The use of scanning ion conductance microscopy to image A6 cells
复制标题
DOI：
10.1016/j.mce.2003.10.015
复制
发表时间：
2004-03-31
影响因子：
4.1
通讯作者：
Korchev, YE
中科院分区：
文献类型：
--
作者：
Gorelik, J;Zhang, YJ;Korchev, YE
回来。-round：对A6活细胞进行连续的高空间分辨率观察，将极大地有助于阐明结构与功能之间的关系，促进醛固酮作用机制等主要生理过程的研究。不幸的是，观察活细胞膜的微观结构和功能变化对显微镜学家来说仍然是一个巨大的挑战。方法：扫描离子电导显微镜（SICM），使用玻璃纳米管作为敏感探针，已被证明适用于成像非导电！表面浸泡在电解质中。我们小组开发了这种显微镜的专门版本，并首次应用于高分辨率活细胞成像。这种方法也可以与贴片夹紧结合使用，以研究解剖和功能，并识别单细胞中的离子通道。结果：这种新显微镜提供了高分辨率的活肾细胞图像，与扫描电子显微镜（SEM）和原子力显微镜（AFM）获得的图像相当。在正常生理条件下的连续24it观察显示A6肾上皮细胞如何改变其高度、体积和重塑其边界。细胞面积的变化与表面微绒毛的密度有关。表面微绒毛密度从扩大细胞的0.5 mum（-2）到缩小细胞的2.5 mum(2)不等。单个细胞的贴片夹紧使解剖和功能相关联。结论：扫描离子电导显微镜提供了关于活细胞的独特信息，有助于了解细胞功能。它有可能成为研究活肾细胞的有力工具。2003爱思唯尔爱尔兰有限公司版权所有。
Back.-round: Continuous high spatial resolution observations of living A6 cells would greatly aid the elucidation of the relationship between structure and function and facilitate the study of major physiological processes Such as the mechanism of action of aldosterone. Unfortunately, observing the micro-structural and functional changes in the membrane of living cells is still a formidable challenge for a microscopist. Method: Scanning ion conductance microscopy (SICM), which uses a glass nanopipette as a sensitive probe, has been shown to be suitable for imaging non-conducting! surfaces bathed in electrolytes. A specialized version of this microscopy has been developed by our group and has been applied to image live cells at high-resolution for the first time. This method can also be used in conjunction with patch clamping to study both anatomy and function and identify ion channels in single cells. Results: This new microscopy provides high-resolution images of living renal cells which are comparable with those obtained by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Continuous 24 It observations under normal physiological conditions showed how A6 kidney epithelial cells changed their height, volume, and reshaped their borders. The changes in cell area correlated with the density of microvilli on the surface. Surface microvilli density ranged from 0.5 mum(-2) for extended cells to 2.5 mum(2) for shrunk cells. Patch clamping of individual cells enabled anatomy and function to be correlated. Conclusions: Scanning ion conductance microscopy provides unique information about living cells that helps to understand cellular function. It has the potential to become a powerful tool for research on living renal cells. (C) 2003 Elsevier Ireland Ltd. All rights reserved.