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用于受激发射损耗显微镜中借助空间光调制器实现轻松系统对准和基本像差传感的二元相位掩膜。

Binary phase masks for easy system alignment and basic aberration sensing with spatial light modulators in STED microscopy.

作者信息

Klauss André, Conrad Florian, Hille Carsten

机构信息

University of Potsdam, Institute of Chemistry, Potsdam, D-14476, Germany.

出版信息

Sci Rep. 2017 Nov 16;7(1):15699. doi: 10.1038/s41598-017-15967-5.

Abstract

The use of binary phase patterns to improve the integration and optimization of spatial light modulators (SLM) in an imaging system, especially a confocal microscope, is proposed and demonstrated. The phase masks were designed to create point spread functions (PSF), which exhibit specific sensitivity to major disturbances in the optical system. This allows direct evaluation of misalignment and fundamental aberration modes by simple visual inspection of the focal intensity distribution or by monitoring the central intensity of the PSF. The use of proposed phase masks is investigated in mathematical modelling and experiment for the use in a stimulated emission depletion (STED) microscope applying wavefront shaping by a SLM. We demonstrate the applicability of these phase masks for modal wavefront sensing of low order aberration modes up to the third order of Zernike polynomials, utilizing the point detector of a confocal microscope in a 'guide star' approach. A lateral resolution of ~25 nm is shown in STED imaging of the confocal microscope retrofitted with a SLM and a STED laser and binary phase mask based system optimization.

摘要

提出并演示了使用二元相位图案来改善成像系统,特别是共聚焦显微镜中空间光调制器(SLM)的集成和优化。相位掩模被设计用于创建点扩散函数(PSF),其对光学系统中的主要干扰表现出特定的敏感性。这允许通过对焦强度分布的简单目视检查或通过监测PSF的中心强度来直接评估对准误差和基本像差模式。在数学建模和实验中研究了所提出的相位掩模在应用SLM进行波前整形的受激发射损耗(STED)显微镜中的应用。我们展示了这些相位掩模对于高达泽尼克多项式三阶的低阶像差模式的模态波前传感的适用性,采用“导星”方法利用共聚焦显微镜的点探测器。在配备有SLM、STED激光器和基于二元相位掩模的系统优化的共聚焦显微镜的STED成像中显示了约25 nm的横向分辨率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1863/5691043/2de2087e4dd2/41598_2017_15967_Fig1_HTML.jpg

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