Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy.
Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy.
复制标题
DOI：
10.1038/s41467-021-24901-3
复制
发表时间：
2021-07-27
影响因子：
16.6
通讯作者：
Fantner GE
中科院分区：
文献类型：
--
作者：
Navikas V;Leitao SM;Grussmayer KS;Descloux A;Drake B;Yserentant K;Werther P;Herten DP;Wombacher R;Radenovic A;Fantner GE
高分辨率活细胞成像对于研究复杂的生物现象是必要的。现代荧光显微镜方法越来越多地与互补的无标记技术相结合，将荧光信息放入细胞环境中。与荧光成像结合使用的最常见的高分辨率成像方法是电子显微镜和原子力显微镜（AFM），最初是为固态材料表征而开发的。原子力显微镜通常可以解析原子台阶，但在软生物样本上，尖端和样本之间的力会使脆弱的膜变形，从而扭曲该技术原本高的轴向分辨率。在这里，我们提出了扫描离子电导显微镜（SICM）作为一种替代方法的地形成像的软生物样品，保持高轴向分辨率的细胞。SICM与活细胞兼容的超分辨率光学波动成像（SOFI）相辅相成。为了证明我们的方法的能力，我们显示了相关的三维细胞图与SOFI实现在二维和三维与自闪烁染料的双色高阶SOFI成像。最后，我们采用相关的SICM/SOFI显微镜可视化肌动蛋白动态在活COS-7细胞亚衍射分辨率。用于对3D细胞表面进行成像的方法通常需要物理相互作用。在这里，作者报告的扫描离子电导显微镜（SICM）和活细胞超分辨率光学起伏成像（SOFI）的软生物样品的非侵入性地形成像相结合。
High-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. Methods for imaging the 3D cell surface often require physical interaction. Here the authors report the combination of scanning ion conductance microscopy (SICM) and live-cell super-resolution optical fluctuation imaging (SOFI) for the non-invasive topographical imaging of soft biological samples.
登录
查看更多内容
影响因子：
2.2
作者：
Berdyyeva, T;Woodworth, CD;Sokolov, I
通讯作者：
Sokolov, I
影响因子：
4.6
作者：
Gomez-Varela, Ana I.;Stamov, Dimitar R.;De Beule, Pieter A. A.
通讯作者：
De Beule, Pieter A. A.
DOI：
10.1073/pnas.0907866106
发表时间：
2009-12-29
影响因子：
11.1
作者：
Dertinger, T.;Colyer, R.;Enderlein, J.
通讯作者：
Enderlein, J.
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：
13.6
作者：
Cosentino, Michela;Canale, Claudio;Diaspro, Alberto
通讯作者：
Diaspro, Alberto