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The use of scanning ion conductance microscopy to image A6 cells

Lookup NU author(s): Professor Chris Edwards

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Abstract

Background: 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 h 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 μm-2 for extended cells to 2.5 μm2 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. © 2003 Elsevier Ireland Ltd. All rights reserved.


Publication metadata

Author(s): Gorelik J, Zhang Y, Shevchuk AI, Frolenkov GI, Sanchez D, Lab MJ, Vodyanoy I, Edwards CRW, Klenerman D, Korchev YE

Editor(s): Vinson, GP and Coghlan, JP

Publication type: Conference Proceedings (inc. Abstract)

Publication status: Published

Conference Name: International Symposium on Aldosterone

Year of Conference: 2004

Pages: 101-108

ISSN: 0303-7207

Publisher: Molecular and Cellular Endocrinology: Elsevier Ireland

URL: http://dx.doi.org/10.1016/j.mce.2003.10.015

DOI: 10.1016/j.mce.2003.10.015

PubMed id: 15134807


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