Scanning Ion Conductance Microscopy for High-Resolution Topography of Soft Samples Including Live Cells

2011 ◽  
Vol 17 (S2) ◽  
pp. 236-237
Author(s):  
G De Filippi ◽  
C Moore
Keyword(s):  
High Resolution ◽  
Live Cells ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy ◽  
Soft Samples

Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.

Development of a Novel Combined Scanning Electrochemical Microscope (SECM) and Scanning Ion-Conductance Microscope (SICM) Probe for Soft Sample Imaging

2012 ◽  
Vol 1422 ◽  
Author(s):  
Andrew J. Pollard ◽  
Nilofar Faruqui ◽  
Michael Shaw ◽  
Charles A. Clifford ◽  
Yasufumi Takahashi ◽  
...  
Keyword(s):  
Focused Ion Beam ◽  
Ion Beam ◽  
Heating System ◽  
Production Method ◽  
Live Cells ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy ◽  
Human Body Temperature ◽  
Variation Versus ◽  
Soft Samples

ABSTRACTBy incorporating a localized heating system within a scanning ion-conductance microscopy (SICM) system, we have performed stable ‘hopping-mode’ (HPICM) imaging for live cells maintained at temperatures ranging up to human body temperature. This allows the accurate study of cell volume and morphology variation versus temperature over extended periods of time. The integration of SICM with scanning electrochemical microscopy (SECM) provides the simultaneous mapping of electrochemical and topographic information for soft samples, such as live cells. This combined technique overcomes the limitations of resolution and topographical artifacts typically associated with SECM. However, previously reported SECM-SICM probe production required expensive and time-consuming focused ion beam (FIB) methods and produced pipettes that are typically hundreds of nanometers in diameter. We report a simple and rapid production method for SECM-SICM double-barrel probes with apertures down to 20 nm in diameter. The characterization of these SECM-SICM probes using scanning electron microscopy (SEM) imaging, cyclic voltammetry (CV) and Raman spectroscopy is also detailed. These SECM-SICM probes were subsequently used to study the morphology and electrochemical activity of several samples, ranging from hard metallic/insulating samples to live cells.

Gauging surface charge distribution of live cell membrane by ionic current change using scanning ion conductance microscopy

Nanoscale ◽  
2021 ◽  
Author(s):  
Feng Chen ◽  
Jin He ◽  
Prakash Manandhar ◽  
Yizi Yang ◽  
Peidang Liu ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Surface Charge ◽  
Charge Distribution ◽  
Ionic Current ◽  
Live Cells ◽  
Ion Conductance ◽  
Physiological Conditions ◽  
Scanning Ion Conductance Microscopy ◽  
Surface Charge Distribution ◽  
Current Change

The distribution of surface charge and potential of cell membrane plays an indispensable role in cellular activities. However, probing surface charge of live cells in physiological conditions, until recently, remains...

Topographic imaging of convoluted surface of live cells by scanning ion conductance microscopy in a standing approach mode

2010 ◽  
Vol 12 (34) ◽  
pp. 10012 ◽  
Author(s):  
Yasufumi Takahashi ◽  
Yumi Murakami ◽  
Kuniaki Nagamine ◽  
Hitoshi Shiku ◽  
Shigeo Aoyagi ◽  
...  
Keyword(s):  
Live Cells ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy ◽  
Topographic Imaging

Imaging Proteins in Membranes of Living Cells by High-Resolution Scanning Ion Conductance Microscopy

2006 ◽  
Vol 45 (14) ◽  
pp. 2212-2216 ◽  
Author(s):  
Andrew I. Shevchuk ◽  
Gregory I. Frolenkov ◽  
Daniel Sánchez ◽  
Peter S. James ◽  
Noah Freedman ◽  
...  
Keyword(s):  
High Resolution ◽  
Living Cells ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy

scholarly journals Imaging the cell surface and its organization down to the level of single molecules

2013 ◽  
Vol 368 (1611) ◽  
pp. 20120027 ◽  
Author(s):  
David Klenerman ◽  
Andrew Shevchuk ◽  
Pavel Novak ◽  
Yuri E. Korchev ◽  
Simon J. Davis
Keyword(s):  
Single Molecule ◽  
Cell Biology ◽  
Two Dimensions ◽  
Live Cells ◽  
High Temporal Resolution ◽  
Biological Processes ◽  
Single Molecule Fluorescence ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy ◽  
And Function

Determining the organization of key molecules on the surface of live cells in two dimensions and how this changes during biological processes, such as signalling, is a major challenge in cell biology and requires methods with nanoscale spatial resolution and high temporal resolution. Here, we review biophysical tools, based on scanning ion conductance microscopy and single-molecule fluorescence and the combination of both of these methods, which have recently been developed to address these issues. We then give examples of how these methods have been be applied to provide new insights into cell membrane organization and function, and discuss some of the issues that will need to be addressed to further exploit these methods in the future.

scholarly journals Scanning ion conductance microscopy: a nanotechnology for biological studies in live cells

2013 ◽  
Vol 3 ◽  
Author(s):  
Bing-Chen Liu ◽  
Xiao-Yu Lu ◽  
Xiang Song ◽  
Ke-Yu Lei ◽  
Abdel A. Alli ◽  
...  
Keyword(s):  
Live Cells ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy ◽  
Biological Studies

scholarly journals Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Vytautas Navikas ◽  
Samuel M. Leitao ◽  
Kristin S. Grussmayer ◽  
Adrien Descloux ◽  
Barney Drake ◽  
...  
Keyword(s):  
High Resolution ◽  
Biological Samples ◽  
Single Cells ◽  
Super Resolution ◽  
Live Cell ◽  
Actin Dynamics ◽  
Label Free ◽  
Axial Resolution ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy

AbstractHigh-resolution live-cell imaging is necessary to study complex biological phenomena. Modern fluorescence microscopy methods are increasingly combined with complementary, label-free techniques to put the fluorescence information into the cellular context. The most common high-resolution imaging approaches used in combination with fluorescence imaging are electron microscopy and atomic-force microscopy (AFM), originally developed for solid-state material characterization. AFM routinely resolves atomic steps, however on soft biological samples, the forces between the tip and the sample deform the fragile membrane, thereby distorting the otherwise high axial resolution of the technique. Here we present scanning ion-conductance microscopy (SICM) as an alternative approach for topographical imaging of soft biological samples, preserving high axial resolution on cells. SICM is complemented with live-cell compatible super-resolution optical fluctuation imaging (SOFI). To demonstrate the capabilities of our method we show correlative 3D cellular maps with SOFI implementation in both 2D and 3D with self-blinking dyes for two-color high-order SOFI imaging. Finally, we employ correlative SICM/SOFI microscopy for visualizing actin dynamics in live COS-7 cells with subdiffraction-resolution.

scholarly journals Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy

2020 ◽  
Author(s):  
Vytautas Navikas ◽  
Samuel M. Leitao ◽  
Kristin S. Grussmayer ◽  
Adrien Descloux ◽  
Barney Drake ◽  
...  
Keyword(s):  
High Resolution ◽  
Biological Samples ◽  
Single Cells ◽  
Super Resolution ◽  
Live Cell ◽  
Actin Dynamics ◽  
Label Free ◽  
Axial Resolution ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy

AbstractHigh-resolution live-cell imaging is necessary to study complex biological phenomena. Modern fluorescence microscopy methods are increasingly combined with complementary, label-free techniques to put the fluorescence information into the cellular context. The most common high-resolution imaging approaches used in combination with fluorescence imaging are electron microscopy and atomic-force microscopy (AFM), originally developed for solid-state material characterization. AFM routinely resolves atomic steps, however on soft biological samples, the forces between the tip and the sample deform the fragile membrane, thereby distorting the otherwise high axial resolution of the technique. Here we present scanning ion-conductance microscopy (SICM) as an alternative approach for topographical imaging of soft biological samples, preserving high axial resolution on cells. SICM is complemented with live-cell compatible super-resolution optical fluctuation imaging (SOFI). To demonstrate the capabilities of our method we show correlative 3D cellular maps with SOFI implementation in both 2D and 3D with self-blinking dyes for two-color high-order SOFI imaging. Finally, we employ correlative SICM/SOFI microscopy for visualizing actin dynamics in live COS-7 cells with subdiffractional resolution.

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