The use of scanning ion conductance microscopy to image A6 cells.
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.


METHODS
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 24h 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 microm(-2) for extended cells to 2.5 microm(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.
Gorelik
Julia
J
Division of Medicine, Imperial College London, MRC Clinical Sciences Centre, Hammersmith Hospital Campus, Du Cane Road, London W12 0NN, UK.
Zhang
Yanjun
Y
Shevchuk
Andrew I
AI
Frolenkov
Gregory I
GI
Sánchez
Daniel
D
Lab
Max J
MJ
Vodyanoy
Igor
I
Edwards
Christopher R W
CR
Klenerman
David
D
Korchev
Yuri E
YE
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure*; D007473:Ion Channels / Q000378:metabolism / Q000648:ultrastructure*; D016252:Microscopy, Scanning Tunneling / Q000379:methods*; D018408:Patch-Clamp Techniques / Q000379:methods*; D014982:Xenopus laevis
D007473:Ion Channels