The use of scanning ion conductance microscopy to image A6 cells (original) (raw)
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Scanning ion conductance microscopy of living cells
Biophysical Journal, 1997
Currently there is a great interest in using scanning probe microscopy to study living cells. However, in most cases the contact the probe makes with the soft surface of the cell deforms or damages it. Here we report a scanning ion conductance microscope specially developed for imaging living cells. A key feature of the instrument is its scanning algorithm, which maintains the working distance between the probe and the sample such that they do not make direct physical contact with each other. Numerical simulation of the probe/sample interaction, which closely matches the experimental observations, provides the optimum working distance. The microscope scans highly convoluted surface structures without damaging them and reveals the true topography of cell surfaces. The images resemble those produced by scanning electron microscopy, with the significant difference that the cells remain viable and active. The instrument can monitor small-scale dynamics of cell surfaces as well as whole-cell movement.
The scanning ion conductance microscope for cellular physiology
AJP: Heart and Circulatory Physiology, 2013
The quest for nonoptical imaging methods that can surmount light diffraction limits resulted in the development of scanning probe microscopes. However, most of the existing methods are not quite suitable for studying biological samples. The scanning ion conductance microscope (SICM) bridges the gap between the resolution capabilities of atomic force microscope and scanning electron microscope and functional capabilities of conventional light microscope. A nanopipette mounted on a three-axis piezo-actuator, scans a sample of interest and ion current is measured between the pipette tip and the sample. The feedback control system always keeps a certain distance between the sample and the pipette so the pipette never touches the sample. At the same time pipette movement is recorded and this generates a three-dimensional topographical image of the sample surface. SICM represents an alternative to conventional high-resolution microscopy, especially in imaging topography of live biological...
Scanning ion conductance microscopy: a nanotechnology for biological studies in live cells
Frontiers in physiology, 2012
Scanning ion-conductance microscope (SICM), which enables high-resolution imaging of cell surface topography, has been developed for over two decades. However, only recently, a unique scanning mode is increasingly used in biological studies to allow SICM to detect the surface of live cells. More recently, in combination with confocal microscopy and patch-clamp electrophysiological techniques, SICM allows investigators to localize proteins or ion channels in a specific nanostructure at the cell surface. This article will briefly review SICM nanotechnique and summarize the role of SICM in biological studies.
Kidney International, 2005
Background. To function as a transport barrier a renal tubule epithelial monolayer needs to maintain its integrity when, sudden hypertonic stress causes cell shrinkage, new cells are added, or cells in the monolayer die. However, the mechanism used to achieve this is largely unknown. Scanning ion conductance microscopy (SICM) has been shown to be suitable for imaging the surface of live renal cells with high topographic resolution, and can be used to elucidate how a functional renal epithelial monolayer maintains its integrity.
Cell Volume Measurement Using Scanning Ion Conductance Microscopy
Biophysical Journal, 2000
We report a novel scanning ion conductance microscopy (SICM) technique for assessing the volume of living cells, which allows quantitative, high-resolution characterization of dynamic changes in cell volume while retaining the cell functionality. The technique can measure a wide range of volumes from 10 Ϫ19 to 10 Ϫ9 liter. The cell volume, as well as the volume of small cellular structures such as lamelopodia, dendrites, processes, or microvilli, can be measured with the 2.5 ϫ 10 Ϫ20 liter resolution. The sample does not require any preliminary preparation before cell volume measurement. Both cell volume and surface characteristics can be simultaneously and continuously assessed during relatively long experiments. The SICM method can also be used for rapid estimation of the changes in cell volume. These are important when monitoring the cell responses to different physiological stimuli.
2011
published online 16 February 2011 J. R. Soc. Interface Gorelik Mitchell, Adrian H. Chester, David Klenerman, Max J. Lab, Yuri E. Korchev, Sian E. Harding and Julia El-Hamamsy, Claire M. F. Potter, Peter Wright, S.H. Sheikh Abdul Kadir, Alexander R. Lyon, Jane A. Michele Miragoli, Alexey Moshkov, Pavel Novak, Andrew Shevchuk, Viacheslav O. Nikolaev, Ismail living cardiovascular cells high-resolution technology for multi-parametric analysis of Scanning ion conductance microscopy: a convergent
Biophysical Journal, 1996
We have constructed a combined TappingMode atomic force microscope and scanning ion conductance microscope. The design is based on a bent glass pipette that acts as both the force sensor and conductance probe. Measuring the pipette deflection allows more stable feedback than possible with previous versions of the scanning ion conductance microscope. Using this microscope, we have imaged synthetic membranes in both contact and tapping modes under fluid. Although contact mode operation is possible, we found that our microscope provided higher contrast and less apparent sample damage in the topographic and ionic conductance images in the tapping mode.