Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy

被引：2074

作者：

Huang, Bo ^{[1
,2
]}

Wang, Wenqin ^{[3
]}

Bates, Mark ^{[4
]}

Zhuang, Xiaowei ^{[1
,2
,3
]}

机构：

[1] Harvard Univ, Howard Hughes Med Inst, Cambridge, MA 02138 USA

[2] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA

[3] Harvard Univ, Dept Phys, Cambridge, MA 02138 USA

[4] Harvard Univ, Sch Engn & Appl Sci, Cambridge, MA 02138 USA

来源：

SCIENCE | 2008年 / 319卷 / 5864期

关键词：

D O I：

10.1126/science.1153529

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：810 / 813

页数：4