Wang Xiao, Zhang Yaxin, Zhou Wenxia, Xu Dongdong, Yin Jianhua
J Opt Soc Am A Opt Image Sci Vis. 2020 Mar 1;37(3):353-360. doi: 10.1364/JOSAA.380805.
Conventional fluorescence polarization microscopy has been largely used to monitor the orientation and the structural information of biomolecules labeled with fluorescence dipoles but suffers from the optical diffraction limit. Here, we put forward a novel algorithm to simultaneously acquire the super-resolution image and the effective orientation distribution information of dipole clusters at corresponding super-resolution. In this paper, the orientation distribution of dipole clusters is statistically modeled by its mean orientation and orientation deviation, which are, respectively, represented by the middle direction and the opening angle of a sector shape. According to this model and microscopy imaging theory, the joint reconstruction algorithm is deduced mathematically in detail based on the conjugate gradient least-squares method. By applying this algorithm to different samples, the reconstructed results prove more than twice the resolution of wide-field images and the orientation distribution information at corresponding spatial resolution. Furthermore, the high accuracy of this algorithm in reconstructing super-resolution orientation distribution information is verified by Monte Carlo simulations.
传统的荧光偏振显微镜在很大程度上已被用于监测用荧光偶极子标记的生物分子的取向和结构信息，但受光学衍射极限的限制。在此，我们提出了一种新颖的算法，能够在相应的超分辨率下同时获取偶极子簇的超分辨率图像和有效取向分布信息。在本文中，偶极子簇的取向分布通过其平均取向和取向偏差进行统计建模，分别由扇形的中间方向和开口角度表示。根据该模型和显微镜成像理论，基于共轭梯度最小二乘法详细推导了联合重建算法。将该算法应用于不同样本，重建结果显示分辨率比宽场图像提高了两倍以上，且在相应空间分辨率下获得了取向分布信息。此外，通过蒙特卡罗模拟验证了该算法在重建超分辨率取向分布信息方面的高精度。
