结构光照明显微镜伪影由照明散射引起。
Structured illumination microscopy artefacts caused by illumination scattering.
机构信息
State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, School of Future Technology, Peking University, Beijing 100871, People's Republic of China.
Center for Bioinformatics, National Laboratory of Protein Engineering and Plant Genetic Engineering, School of Life Sciences, Peking University, Beijing 100871, People's Republic of China.
出版信息
Philos Trans A Math Phys Eng Sci. 2021 Jun 14;379(2199):20200153. doi: 10.1098/rsta.2020.0153. Epub 2021 Apr 26.
Despite its wide application in live-cell super-resolution (SR) imaging, structured illumination microscopy (SIM) suffers from aberrations caused by various sources. Although artefacts generated from inaccurate reconstruction parameter estimation and noise amplification can be minimized, aberrations due to the scattering of excitation light on samples have rarely been investigated. In this paper, by simulating multiple subcellular structure with the distinct refractive index from water, we study how different thicknesses of this subcellular structure scatter incident light on its optical path of SIM excitation. Because aberrant interference light aggravates with the increase in sample thickness, the reconstruction of the 2D-SIM SR image degraded with the change of focus along the axial axis. Therefore, this work may guide the future development of algorithms to suppress SIM artefacts caused by scattering in thick samples. This article is part of the Theo Murphy meeting issue 'Super-resolution structured illumination microscopy (part 1)'.
尽管结构光照明显微镜(SIM)在活细胞超分辨率(SR)成像中得到了广泛的应用,但它仍受到各种来源的像差的影响。尽管可以最小化由于不准确的重建参数估计和噪声放大产生的伪影,但由于激发光在样品上散射而产生的像差很少被研究。在本文中,通过模拟具有与水不同折射率的多个亚细胞结构,我们研究了不同厚度的亚细胞结构如何散射入射光在 SIM 激发的光路上。由于异常干涉光随样品厚度的增加而加剧,因此 2D-SIM SR 图像的重建随轴向焦点的变化而退化。因此,这项工作可能指导未来开发抑制厚样品散射引起的 SIM 伪影的算法。本文是 Theo Murphy 会议议题“超分辨率结构光照明显微镜(第 1 部分)”的一部分。