Acta Phys. -Chim. Sin. ›› 2016, Vol. 32 ›› Issue (5): 1123-1128.doi: 10.3866/PKU.WHXB201603234

• ARTICLE • Previous Articles     Next Articles

Super-Resolution Optical Subtraction Microscopy Using Optical Scattering Imaging

Qian ZHOU1,2,3,Jian-Qiang YU4,Li-Bo ZHAO3,De-Sheng LI5,Kui WU6,Jian-Hua ZHU1,2,Jing-He YUAN3,*(),Xiao-Hong FANG3   

  1. 1 Department of Physics, Sichuan University, Chengdu 610064, P. R. China
    2 Key Laboratory of High Energy Density Physics and Technology of Ministry of Education, Chengdu 610064, P. R. China
    3 Key Laboratory of Molecular Nanostructure and Nanotechnology, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, P. R. China
    4 College of Science, North China University of Technology, Beijing 100144, P. R. China
    5 State Key Laboratory of Polymer Physics and Chemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, P. R. China
    6 Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, P. R. China
  • Received:2016-02-25 Published:2016-05-07
  • Contact: Jing-He YUAN E-mail:jhyuan@iccas.ac.cn
  • Supported by:
    the National Key Basic Research Program of China (973)(2013CB933701);National Natural Science Foundation of China(21127901);National Natural Science Foundation of China(91413119);Key Technology Talent Program of Chinese Academy of Sciences

Abstract:

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.

Key words: Super-resolution microscopy, Subtraction imaging, Non-fluorescence imaging, Optical scattering imaging

MSC2000: 

  • O644