Hopping Probe Ion Conductance Microscopy and its Combined Patch-Clamping:A Powerful Tool for Structural and Functional Studies in Neural Nanobiology

2011 ◽  
Vol 694 ◽  
pp. 54-58
Author(s):  
Xin Liang Zhao ◽  
Xiao Liu ◽  
Hu Jie Lu ◽  
Li Ying Ma ◽  
Rui Ling Gao ◽  
...  

Continuous high-resolution observations of cell membrane would greatly aid the elucidation of the relationship between structure and function and facilitate the study of physiological processing in cell biology. However, high-resolution studying living neuron membrane structures and its functions is still a challenge in current nanobiology. The new developed Hoping Probe Ion Conductance Microscopy (HPICM) is designed for non-contact continuous high-resolution topographic imaging of living cells under physiological conditions. In this review, we concisely introduced the basic operation principle of HPICM and its applications in high spatial resolution imaging of two living neuron cell models, N-type SK-N-SH cells and NGF-differentiated sympathetic neuron-like PC12 cells. Combining HPICM with patch-clamp technique, we further investigated the functional ion-channel of under-differentiated neuron-like PC12 cells and demonstrated that NGF treatment promoted the outgrowth of neurites and increased the activity of TTX-sensitive sodium channel. All these results demonstrate that HPICM combined with patch-clamp technique offers high-resolution topographic imaging of living neurons with non-contact — making HPICM an ideal high-resolution imaging technique not to interact/interfere with living neurons during image acquisition, and provides detailed information about the relationship between membrane structures and ion-channel functions of living neurons at the same time, which has the potential to become a powerful microscopy for in-depth researching in neural nanobiology.

1991 ◽  
Vol 261 (5) ◽  
pp. F808-F814 ◽  
Author(s):  
H. Matsunaga ◽  
N. Yamashita ◽  
Y. Miyajima ◽  
T. Okuda ◽  
H. Chang ◽  
...  

We used the patch-clamp technique to clarify the nature of ion channels in renal mesangial cells in culture. In the cell-attached mode most patches were silent in the absence of agonists. In some patches a 25-pS nonselective channel was observed. This 25-pS cation channel was consistently observed in inside-out patches, and it was activated by intracellular Ca2+. Excised patch experiments also revealed the existence of a 40-pS K+ channel, which was activated by intracellular Ca2+. This 40-pS K+ channel was observed infrequently in the cell-attached mode. The activities of both channels were increased by arginine vasopressin or angiotensin II, resulting from an increase in intracellular Ca2+ concentration.


1996 ◽  
Vol 07 (04) ◽  
pp. 321-331 ◽  
Author(s):  
LARRY S. LIEBOVITCH ◽  
ANGELO T. TODOROV

Ion channels in the cell membrane spontaneously switch from states that are closed to the flow of ions such as sodium, potassium, and chloride to states that are open to the flow of these ions. The durations of times that an individual ion channel protein spends in the closed and open states can be measured by the patch clamp technique. We explore two basic issues about the molecular properties of ion channels: 1) If the switching between the closed and open state is an inherently random event, what does the patch clamp data tell us about the structure or motions in the ion channel protein? 2) Is this switching random?


2013 ◽  
Vol 446-447 ◽  
pp. 427-433 ◽  
Author(s):  
Xiao Lin He ◽  
Jin Hai Wang ◽  
Yu Zheng ◽  
Yang Zhang ◽  
Gang Li

In order to get the single cortical neurons fitting the study of patch-clamp technique.use the kunming mouse with the birth of 10 days or so, and separate neurons in the cerebral cortex through acute separation technology and enzymatic hydrolysis. A cone-shaped adherent neurons are selected and observed in rapid dissociation and cell activity status under the inverted microscope. Different ion channel blockers is added to the extracellular fluid and used whole-cell recording which can obtain Na +, K + and other ion channel currents. This method can obtain good morphological and physiological characteristics of both single cortical neurons, confirming that the method applied to cortical neurons by patch clamp technique, which has significant value for in-depth research on drugs, toxins and other physical factors on the electrical activity of neuronal ion channels and signal transduction mechanism of the effect.


Physiology ◽  
1990 ◽  
Vol 5 (4) ◽  
pp. 155-158
Author(s):  
C Zuazaga ◽  
A Steinacker

Glass for pipettes used to record ion channel activity with the patch-clamp technique is selected on the basis of its electrical, thermal, and sealing properties. Recent findings stress a new characteristic to consider: the effect of pipette glass itself on ion channel properties.


2017 ◽  
Vol 132 ◽  
pp. 45-52 ◽  
Author(s):  
Rigmor C. Baraas ◽  
Jon V.B. Gjelle ◽  
Elisabeth Bratlie Finstad ◽  
Siri Bjørnetun Jacobsen ◽  
Stuart J. Gilson

2004 ◽  
Vol 5 (4) ◽  
pp. 387-395 ◽  
Author(s):  
K. Jurkat-Rott ◽  
F. Lehmann-Horn

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