Low Stress Ion Conductance Microscopy of Sub-Cellular Stiffness.
Low Stress Ion Conductance Microscopy of Sub-Cellular Stiffness.
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DOI：
10.1039/c6sm01106c
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发表时间：
2016-10-14
期刊：
影响因子：
3.4
通讯作者：
Klenerman D
中科院分区：
文献类型：
--
作者：
Clarke RW;Novak P;Zhukov A;Tyler EJ;Cano-Jaimez M;Drews A;Richards O;Volynski K;Bishop C;Klenerman D
量化离子电导显微镜固有的力使其能够映射亚细胞结构的刚度，即使非常软。直接检查亚细胞力学，同时避免活细胞的过度应变，需要精确控制非常小区域的光应力，这从根本上是困难的。在这里，我们使用一个不与质膜接触的玻璃纳米吸管，既对细胞施加压力，又准确地监测细胞压缩。这允许在比100 nm更精细的横向分辨率下绘制细胞刚度。我们将纳米移液管施加在细胞上的应力计算为尖端面和质膜之间的固有压力加上其对任何糖萼的直接压力的总和，两者均根据离子电流减小的差距大小进行评估。对细胞类型的研究证实，大约120 Pa的细胞内压力开始使质膜与细胞骨架分离，并揭示神经元细胞体的第一个0.66 ± 0.09 μm的压缩比以前的方法能够检测到的要软得多。
Quantifying forces inherent to ion conductance microscopy enables it to map the stiffness of sub-cellular structures, even if very soft. Directly examining subcellular mechanics whilst avoiding excessive strain of a live cell requires the precise control of light stress on very small areas, which is fundamentally difficult. Here we use a glass nanopipet out of contact with the plasma membrane to both exert the stress on the cell and also accurately monitor cellular compression. This allows the mapping of cell stiffness at a lateral resolution finer than 100 nm. We calculate the stress a nanopipet exerts on a cell as the sum of the intrinsic pressure between the tip face and the plasma membrane plus its direct pressure on any glycocalyx, both evaluated from the gap size in terms of the ion current decrease. A survey of cell types confirms that an intracellular pressure of approximately 120 Pa begins to detach the plasma membrane from the cytoskeleton and reveals that the first 0.66 ± 0.09 μm of compression of a neuron cell body is much softer than previous methods have been able to detect.
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影响因子：
16.2
作者：
Novak P;Gorelik J;Vivekananda U;Shevchuk AI;Ermolyuk YS;Bailey RJ;Bushby AJ;Moss GW;Rusakov DA;Klenerman D;Kullmann DM;Volynski KE;Korchev YE
通讯作者：
Korchev YE
DOI：
10.1126/science.1241224
发表时间：
2013-10-18
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Xie L;Kang H;Xu Q;Chen MJ;Liao Y;Thiyagarajan M;O'Donnell J;Christensen DJ;Nicholson C;Iliff JJ;Takano T;Deane R;Nedergaard M
通讯作者：
Nedergaard M
影响因子：
14.9
作者：
Overhoff MG;Garbe JC;Koh J;Stampfer MR;Beach DH;Bishop CL
通讯作者：
Bishop CL
影响因子：
48
作者：
Novak, Pavel;Li, Chao;Shevchuk, Andrew I.;Stepanyan, Ruben;Caldwell, Matthew;Hughes, Simon;Smart, Trevor G.;Gorelik, Julia;Ostanin, Victor P.;Lab, Max J.;Moss, Guy W. J.;Frolenkov, Gregory I.;Klenerman, David;Korchev, Yuri E.
通讯作者：
Korchev, Yuri E.
影响因子：
3.4
作者：
Charras, Guillaume T.;Coughlin, Margaret;Mahadevan, L.
通讯作者：
Mahadevan, L.