Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy

by: Bo Huang, Wenqin Wang, Mark Bates, Xiaowei Zhuang

Science (3 January 2008), 1153529.

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Abstract

Recent advances in far-field fluorescence microscopy have led to substantial improvements in image resolution, achieving a near-molecular resolution of 20 30 nm in the two lateral dimensions. Three-dimensional (3D) nanoscale-resolution imaging, however, remains a challenge. Here, we demonstrate 3D stochastic optical reconstruction microscopy (STORM) by determining both axial and lateral positions of individual fluorophores with nanometer accuracy using optical astigmatism. Iterative, stochastic activation of photo-switchable 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 30 nm in the lateral dimensions and 50 60 nm in the axial dimension. This development allowed us to resolve the 3D morphology of nanoscopic cellular structures. 10.1126/science.1153529

@article{citeulike:2205894, abstract = {Recent advances in far-field fluorescence microscopy have led to substantial improvements in image resolution, achieving a near-molecular resolution of 20 30 nm in the two lateral dimensions. Three-dimensional (3D) nanoscale-resolution imaging, however, remains a challenge. Here, we demonstrate 3D stochastic optical reconstruction microscopy (STORM) by determining both axial and lateral positions of individual fluorophores with nanometer accuracy using optical astigmatism. Iterative, stochastic activation of photo-switchable 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 30 nm in the lateral dimensions and 50 60 nm in the axial dimension. This development allowed us to resolve the 3D morphology of nanoscopic cellular structures. 10.1126/science.1153529}, author = {Huang, Bo and Wang, Wenqin and Bates, Mark and Zhuang, Xiaowei }, citeulike-article-id = {2205894}, doi = {http://dx.doi.org/10.1126/science.1153529}, journal = {Science}, keywords = {cell, imaging}, month = {January}, pages = {1153529+}, posted-at = {2008-02-10 21:17:51}, priority = {2}, title = {Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy}, url = {http://dx.doi.org/10.1126/science.1153529}, year = {2008} }

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TY - JOUR ID - citeulike:2205894 L3 - citeulike-article-id:2205894 TI - Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy JF - Science SP - 1153529 N2 - Recent advances in far-field fluorescence microscopy have led to substantial improvements in image resolution, achieving a near-molecular resolution of 20 30 nm in the two lateral dimensions. Three-dimensional (3D) nanoscale-resolution imaging, however, remains a challenge. Here, we demonstrate 3D stochastic optical reconstruction microscopy (STORM) by determining both axial and lateral positions of individual fluorophores with nanometer accuracy using optical astigmatism. Iterative, stochastic activation of photo-switchable 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 30 nm in the lateral dimensions and 50 60 nm in the axial dimension. This development allowed us to resolve the 3D morphology of nanoscopic cellular structures. 10.1126/science.1153529 KW - cell KW - imaging AU - Huang, Bo AU - Wang, Wenqin AU - Bates, Mark AU - Zhuang, Xiaowei PY - 2008/01/03/ UR - http://dx.doi.org/10.1126/science.1153529 ER -

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