Whole-cell 3D STORM reveals interactions between cellular structures with nanometer-scale resolution.
Whole-cell 3D STORM reveals interactions between cellular structures with nanometer-scale resolution.
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DOI：
10.1038/nmeth.1274
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发表时间：
2008-12
期刊：
影响因子：
48
通讯作者：
Zhuang, Xiaowei
中科院分区：
文献类型：
--
作者：
Huang, Bo;Jones, Sara A.;Brandenburg, Boerries;Zhuang, Xiaowei
直接可视化纳米细胞结构及其三维空间关系的能力将极大地增强我们对细胞分子过程的理解。在这里，我们展示了多色三维（3D）随机光学重建显微镜（STORM）作为定量探测细胞结构及其相互作用的工具。为了促进STORM成像，我们通过共价连接一个可光切换的菁氨酸报告分子和一个激活分子来辅助生物偶联，产生了几种不同颜色的可光切换探针。三维定位结合焦平面扫描和折射率失配校正，获得横向和轴向空间分辨率分别为20-30 nm和60-70 nm的全细胞图像。利用这种方法，我们对固定猴肾BS-C-1细胞的整个线粒体网络进行了成像，并研究了线粒体与微管之间的空间关系。3D STORM图像显示了线粒体形态以及线粒体-微管接触，这些在常规荧光图像中是模糊的。
The ability to directly visualize nanoscopic cellular structures and their spatial relationship in all three dimensions will greatly enhance our understanding of molecular processes in cells. Here, we demonstrated multicolor three-dimensional (3D) stochastic optical reconstruction microscopy (STORM) as a tool to quantitatively probe cellular structures and their interactions. To facilitate STORM imaging, we generated photoswitchable probes in several distinct colors by covalently linking a photoswitchable cyanine reporter and an activator molecule to assist bioconjugation. 3D localization was performed in conjunction with focal plane scanning and correction for refractive index mismatch to obtain whole-cell images with a spatial resolution of 20–30 nm and 60–70 nm in the lateral and axial dimensions, respectively. Using this approach, we imaged the entire mitochondrial network in fixed monkey kidney BS-C-1 cells, and studied the spatial relationship between mitochondria and microtubules. The 3D STORM images revealed mitochondrial morphologies as well as mitochondria-microtubule contacts that were obscured in conventional fluorescence images.
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56.9
作者：
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