A New Method for Axial Decay Function Calibration of Evanescent Field in Multi-Angle Total Internal Reflection Fluorescence Microscopy

5th International Conference on Advances in Optoelectronics and Micro/Nano-optics

Wu, Jian^1,2 ; Xiu, Peng^3 ; Jin, Luhong^1,2 ; Nan, Di^1,2 ; Kuang, Cuifang^3 ; Zheng, Xiaoxiang^1,2,4 ; Xu, Yingke^1,2 ; Liu, Xu^3

Key Laboratory for Biomedical Engineering of Ministry of Education, Department of Biomedical Engineering, Zhejiang University, Hangzhouv, China^1

Zhejiang Prov. Key Lab. of Cardio-Cerebr. Vasc. Detection Technol. and Med. Effectiveness Appraisal, Zhejiang University, Hangzhou

310027, China^2

State Key Laboratory of Modern Optical Instrumentation, Department of Optical Engineering, Zhejiang University, Hangzhou

310027, China^3

Qiushi Academy for Advanced Studies, Zhejiang University, Hangzhou

310027, China^4

关键词: 3D reconstruction;    Axial resolutions;    Calibration method;    Evanescent wave;    Incidence angles;    Theoretical equation;    Three-dimensional image reconstruction;    Total internal reflection fluorescence microscopy;   

Others  :  https://iopscience.iop.org/article/10.1088/1742-6596/680/1/012025/pdf
DOI  :  10.1088/1742-6596/680/1/012025

来源: IOP

【 摘 要 】

Three-dimensional image reconstruction in multi-angle total internal reflection fluorescence microscopy relies on actual penetration depths of evanescent wave at various incidence angles. In this paper, we propose a simple and elegant calibration method to calculate the actual axial decay profile of a given evanescent field, for the analytical solution of theoretical equation is hard to solve in complicated conditions. The results calculated by the proposed method agree with the experimental demands in our research. Our calibration method, together with multi-angle TIRF imaging, permits 3D reconstruction of cell surface in superb axial resolution.