Visualizing intracellular nanostructures of living cells by nanoendoscopy-AFM
Visualizing intracellular nanostructures of living cells by nanoendoscopy-AFM
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DOI：
10.1126/sciadv.abj4990
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发表时间：
2021-12-01
期刊：
影响因子：
13.6
通讯作者：
Fukuma, Takeshi
中科院分区：
文献类型：
--
作者：
Penedo, Marcos;Miyazawa, Keisuke;Fukuma, Takeshi
原子力显微镜（AFM）是唯一一种允许对纳米级生物分子动力学进行无标记成像的技术，在解决其他主要生物成像工具（荧光或电子显微镜）无法解决的生物学问题方面发挥着至关重要的作用。然而，这样的成像仅可能用于从细胞提取或在固体基质上重建的系统。因此，活细胞内的纳米动力学在很大程度上仍然无法与目前的纳米成像技术。在这里，我们通过纳米内窥镜-AFM克服了这一限制，其中针状纳米探针插入活细胞中，呈现肌动蛋白纤维三维（3D）图和膜内支架的2D纳米动力学，导致细胞活力的不可检测的变化。与以前的AFM方法不同，纳米探针直接访问目标细胞内成分，利用AFM的所有功能，如高分辨率成像，纳米力学映射和分子识别。这些特征应该大大扩展了在活细胞中可观察到的细胞内结构的范围。
Atomic force microscopy (AFM) is the only technique that allows label-free imaging of nanoscale biomolecular dynamics, playing a crucial role in solving biological questions that cannot be addressed by other major bioimaging tools (fluorescence or electron microscopy). However, such imaging is possible only for systems either extracted from cells or reconstructed on solid substrates. Thus, nanodynamics inside living cells largely remain inaccessible with the current nanoimaging techniques. Here, we overcome this limitation by nanoendoscopy-AFM, where a needle-like nanoprobe is inserted into a living cell, presenting actin fiber three-dimensional (3D) maps, and 2D nanodynamics of the membrane inner scaffold, resulting in undetectable changes in cell viability. Unlike previous AFM methods, the nanoprobe directly accesses the target intracellular components, exploiting all the AFM capabilities, such as high-resolution imaging, nanomechanical mapping, and molecular recognition. These features should greatly expand the range of intracellular structures observable in living cells.