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提高相干结构照明显微镜横向分辨率的各向同性

Enhancing the isotropy of lateral resolution in coherent structured illumination microscopy.

作者信息

Park Joo Hyun, Lee Jae Yong, Lee Eun Seong

机构信息

Nano and Bio Surface Science, University of Science and Technology, Daejeon 305-350, South Korea ; These authors contributed equally to this work.

Nano and Bio Surface Science, University of Science and Technology, Daejeon 305-350, South Korea ; Center for Length, Korea Research Institute of Standards and Science, Daejeon 304-340, South Korea ; These authors contributed equally to this work.

出版信息

Biomed Opt Express. 2014 May 21;5(6):1895-912. doi: 10.1364/BOE.5.001895. eCollection 2014 Jun 1.

Abstract

We present a method to improve the isotropy of spatial resolution in a structured illumination microscopy (SIM) implemented for imaging non-fluorescent samples. To alleviate the problem of anisotropic resolution involved with the previous scheme of coherent SIM that employs the two orthogonal standing-wave illumination, referred to as the orthogonal SIM, we introduce a hexagonal-lattice illumination that incorporates three standing-wave fields simultaneously superimposed at the orientations equally divided in the lateral plane. A theoretical formulation is worked out rigorously for the coherent image formation with such a simultaneous multiple-beam illumination and an explicit Fourier-domain framework is derived for reconstructing an image with enhanced resolution. Using a computer-synthesized resolution target as a 2D coherent sample, we perform numerical simulations to examine the imaging characteristics of our three-angle SIM compared with the orthogonal SIM. The investigation on the 2D resolving power with the various test patterns of different periods and orientations reveal that the orientation-dependent undulation of lateral resolution can be reduced from 27% to 8% by using the three-angle SIM while the best resolution (0.54 times the resolution limit of conventional coherent imaging) in the directions of structured illumination is slightly deteriorated by 4.6% from that of the orthogonal SIM.

摘要

我们提出了一种方法,用于在为非荧光样本成像而实现的结构照明显微镜(SIM)中提高空间分辨率的各向同性。为了缓解与先前采用两个正交驻波照明的相干SIM方案(称为正交SIM)相关的各向异性分辨率问题,我们引入了一种六边形晶格照明,该照明同时包含三个在横向平面上等分方向叠加的驻波场。针对这种同时多光束照明的相干图像形成,我们严格推导了理论公式,并得出了用于重建高分辨率图像的显式傅里叶域框架。使用计算机合成的分辨率目标作为二维相干样本,我们进行了数值模拟,以研究我们的三角度SIM与正交SIM相比的成像特性。对具有不同周期和方向的各种测试图案的二维分辨能力的研究表明,使用三角度SIM时,横向分辨率的方向依赖性波动可从27%降低到8%,而在结构照明方向上的最佳分辨率(传统相干成像分辨率极限的0.54倍)与正交SIM相比略有下降,下降了4.6%。

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