Generation of three-dimensional fluorescent spot  with radially polarized Laguerre-Gaussian beams for STED microscopy imaging

Mingyan Luo; Yanju Yang; Yingshun Xu; Shujing Liu; Dongqing Sun; Shuguang Yang; Zengguang Ma

doi:10.1117/12.2505207

12 December 2018 Generation of three-dimensional fluorescent spot with radially polarized Laguerre-Gaussian beams for STED microscopy imaging

Mingyan Luo, Yanju Yang, Yingshun Xu, Shujing Liu, Dongqing Sun, Shuguang Yang, Zengguang Ma

Author Affiliations +

Proceedings Volume 10846, Optical Sensing and Imaging Technologies and Applications; 108461X (2018) https://doi.org/10.1117/12.2505207
Event: International Symposium on Optoelectronic Technology and Application 2018, 2018, Beijing, China

Abstract

It is very important to have high spatial resolution in all three dimensions for imaging complex 3D biological structures. In this paper, the three-dimensional (3D) fluorescent spot breaking the diffraction-limited resolution barrier is produced by radially polarized Laguerre-Gaussian(LG) beams from stimulated emission depletion(STED) principle. The pump beam is generated by 4π tight focusing of LG₀₁ with high numerical aperture(NA), the full width at half maximum(FWHM) of longitudinal intensity field is 0.42λ , the FWHM of transverse component field is 0.48 λ . The depletion 3D hollow dark spot beam is generated with a topological charge of 1 LG₁₁ through high NA tight focusing. The behavior of the two-beam produced fluorescent spot is examined and the fluorescent spot size is minimized obviously. The super resolutions of 0.034λ and 0.094λ for longitudinal and transverse directions can be achieved, respectively.

Citation Download Citation

Mingyan Luo, Yanju Yang, Yingshun Xu, Shujing Liu, Dongqing Sun, Shuguang Yang, and Zengguang Ma "Generation of three-dimensional fluorescent spot with radially polarized Laguerre-Gaussian beams for STED microscopy imaging", Proc. SPIE 10846, Optical Sensing and Imaging Technologies and Applications, 108461X (12 December 2018); https://doi.org/10.1117/12.2505207

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KEYWORDS

Stimulated emission depletion microscopy

Microscopy

3D image processing

Spherical lenses

Objectives

Super resolution microscopy

Diffraction

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