Time-Resolved Scanning Ion Conductance Microscopy for Three-Dimensional Tracking of Nanoscale Cell Surface Dynamics.
Time-Resolved Scanning Ion Conductance Microscopy for Three-Dimensional Tracking of Nanoscale Cell Surface Dynamics.
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DOI：
10.1021/acsnano.1c05202
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发表时间：
2021-11-23
期刊：
影响因子：
17.1
通讯作者：
Fantner GE
中科院分区：
文献类型：
--
作者：
Leitao SM;Drake B;Pinjusic K;Pierrat X;Navikas V;Nievergelt AP;Brillard C;Djekic D;Radenovic A;Persat A;Constam DB;Anders J;Fantner GE
纳米表征在理解细胞和细胞器的复杂纳米级组织中起着至关重要的作用。了解细胞功能需要关于细胞结构如何随时间演变的高分辨率信息。存在许多技术来非常详细地解析细胞的静态纳米级结构（超分辨率光学显微镜、EM、AFM）。然而，时间分辨成像技术往往具有较低的分辨率，限于小区域，或对细胞造成损伤，从而阻止长期的时间推移研究。诸如原子力显微镜（AFM）的扫描探针显微镜方法将联合收割机高分辨率成像与在生理条件下对活细胞成像的能力相结合。然而，尖端和样品之间的机械接触使细胞表面变形，干扰了天然状态，并阻止了长期的延时成像。在这里，我们开发了一种扫描离子电导显微镜（SICM）的高速和长期的纳米级真核细胞成像。通过利用纳米定位，纳米孔制造，微电子学和控制工程的进步，我们开发了一种显微镜方法，可以解决时空多样的三维（3D）过程中的细胞膜在亚5纳米轴向分辨率。我们用纳米级的细节和亚秒到几天的时间范围跟踪活细胞形态的动态变化，成像不同的过程，从内吞作用，微胞饮作用，有丝分裂到细菌感染和癌细胞的细胞分化。这项技术能够详细观察膜事件，并可能为感染，免疫学和癌症研究提供细胞-细胞相互作用的见解。
Nanocharacterization plays a vital role in understanding the complex nanoscale organization of cells and organelles. Understanding cellular function requires high-resolution information about how the cellular structures evolve over time. A number of techniques exist to resolve static nanoscale structure of cells in great detail (super-resolution optical microscopy, EM, AFM). However, time-resolved imaging techniques tend to either have a lower resolution, are limited to small areas, or cause damage to the cells, thereby preventing long-term time-lapse studies. Scanning probe microscopy methods such as atomic force microscopy (AFM) combine high-resolution imaging with the ability to image living cells in physiological conditions. The mechanical contact between the tip and the sample, however, deforms the cell surface, disturbs the native state, and prohibits long-term time-lapse imaging. Here, we develop a scanning ion conductance microscope (SICM) for high-speed and long-term nanoscale imaging of eukaryotic cells. By utilizing advances in nanopositioning, nanopore fabrication, microelectronics, and controls engineering, we developed a microscopy method that can resolve spatiotemporally diverse three-dimensional (3D) processes on the cell membrane at sub-5-nm axial resolution. We tracked dynamic changes in live cell morphology with nanometer details and temporal ranges of subsecond to days, imaging diverse processes ranging from endocytosis, micropinocytosis, and mitosis to bacterial infection and cell differentiation in cancer cells. This technique enables a detailed look at membrane events and may offer insights into cell–cell interactions for infection, immunology, and cancer research.
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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：
Eghiaian, Frederic;Rigato, Annafrancesca;Scheuring, Simon
通讯作者：
Scheuring, Simon
影响因子：
16.6
作者：
Navikas V;Leitao SM;Grussmayer KS;Descloux A;Drake B;Yserentant K;Werther P;Herten DP;Wombacher R;Radenovic A;Fantner GE
通讯作者：
Fantner GE
影响因子：
16.6
作者：
Klausen, Lasse Hyldgaard;Fuhs, Thomas;Dong, Mingdong
通讯作者：
Dong, Mingdong
影响因子：
64.8
作者：
Johannessen, Cory M.;Johnson, Laura A.;Piccioni, Federica;Townes, Aisha;Frederick, Dennie T.;Donahue, Melanie K.;Narayan, Rajiv;Flaherty, Keith T.;Wargo, Jennifer A.;Root, David E.;Garraway, Levi A.
通讯作者：
Garraway, Levi A.