Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy
Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy
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DOI：
10.1126/science.1153529
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发表时间：
2008-02-08
期刊：
影响因子：
56.9
通讯作者：
Zhuang, Xiaowei
中科院分区：
文献类型：
--
作者：
Huang, Bo;Wang, Wenqin;Zhuang, Xiaowei
远场荧光显微镜的最新进展导致图像分辨率的大幅提高，在两个横向维度上实现了20至30纳米的近分子分辨率。然而，三维(3D)纳米分辨率成像仍然是一个挑战。我们展示了3D随机光学重建显微镜(STORM)，它利用光学散光以纳米精度确定单个荧光团的轴向和横向位置。重复、随机地激活可光切换探头使得能够对每个探头进行高精度的3D定位，从而无需扫描样本即可构建3D图像。使用这种方法，我们在横向维度上获得了20到30纳米的图像分辨率，在轴向维度上获得了50到60纳米的图像分辨率。这一发展使我们能够解析纳米细胞结构的3D形态。
Recent advances in far- field fluorescence microscopy have led to substantial improvements in image resolution, achieving a near- molecular resolution of 20 to 30 nanometers in the two lateral dimensions. Three- dimensional ( 3D) nanoscale- resolution imaging, however, remains a challenge. We demonstrated 3D stochastic optical reconstruction microscopy ( STORM) by using optical astigmatism to determine both axial and lateral positions of individual fluorophores with nanometer accuracy. Iterative, stochastic activation of photoswitchable probes enables high- precision 3D localization of each probe, and thus the construction of a 3D image, without scanning the sample. Using this approach, we achieved an image resolution of 20 to 30 nanometers in the lateral dimensions and 50 to 60 nanometers in the axial dimension. This development allowed us to resolve the 3D morphology of nanoscopic cellular structures.