Citation: ZHOU Qian, YU Jian-Qiang, ZHAO Li-Bo, LI De-Sheng, WU Kui, ZHU Jian-Hua, YUAN Jing-He, FANG Xiao-Hong. Super-Resolution Optical Subtraction Microscopy Using Optical Scattering Imaging[J]. Acta Physico-Chimica Sinica, ;2016, 32(5): 1123-1128. doi: 10.3866/PKU.WHXB201603234 shu

Super-Resolution Optical Subtraction Microscopy Using Optical Scattering Imaging

ZHOU Qian ^1,2,3 ,
YU Jian-Qiang ⁴ ,
ZHAO Li-Bo ³ ,
LI De-Sheng ⁵ ,
WU Kui ⁶ ,
ZHU Jian-Hua ^1,2 ,
YUAN Jing-He ^3,, ,
FANG Xiao-Hong ³

1.
1 Department of Physics, Sichuan University, Chengdu 610064, P. R. China;
2.
2 Key Laboratory of High Energy Density Physics and Technology of Ministry of Education, Chengdu 610064, P. R. China;
3.
3 Key Laboratory of Molecular Nanostructure and Nanotechnology, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, P. R. China;
4.
4 College of Science, North China University of Technology, Beijing 100144, P. R. China;
5.
5 State Key Laboratory of Polymer Physics and Chemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, P. R. China;
6.
6 Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, P. R. China

Corresponding author: YUAN Jing-He,
Received Date: 25 February 2016
Available Online: 22 March 2016
Fund Project: 国家重点基础研究发展规划项目(973)(2013CB933701) (973)(2013CB933701)国家自然科学基金(21127901,91413119) (21127901,91413119)

The existing form of super-resolution microscopy based on specific fluorescent tagging is unable to obtain super-resolution images of non-fluorescent samples. Hence, we have developed optical subtraction microscopy for obtaining super-resolution imaging in such cases. This method is based on image subtraction between the two optical scattering images from general confocal excitation and doughnut-shaped excitation, respectively. Unlike super-resolution fluorescence microscopy, subtraction microscopy requires no preprocessing of the sample, and the excitation power can be kept low to avoid sample damage. The non-fluorescent imaging of gold nanobeads and polymer nanofibers has been realized to demonstrate the feasibility of super-resolution subtraction microscopy. The lateral resolution decreases to 215 nm (0.33λ, 1λ = 650 nm) in subtraction imaging, and greater imaging detail of the sample is achieved via optical scattering.
- Super-resolution microscopy,
- Subtraction imaging,
- Non-fluorescence imaging,
- Optical scattering imaging
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