Experimental assessment and analysis of super-resolution in fluorescence microscopy based on multiple-point spread function fitting of spectrally demultiplexed images
Experimental assessment and analysis of super-resolution in fluorescence microscopy based on multiple-point spread function fitting of spectrally demultiplexed images
复制标题
基于光谱解复用图像多点扩散函数拟合的荧光显微镜超分辨率实验评估和分析
DOI：
10.1007/s10043-017-0379-y
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发表时间：
2018
期刊：
影响因子：
1.2
通讯作者：
Tanida Jun
中科院分区：
文献类型：
--
作者：
Nishimura Takahiro;Kimura Hitoshi;Ogura Yusuke;Tanida Jun
本文提出了一种实验评估和分析的超分辨率显微镜的基础上多点扩散函数拟合的光谱解复用图像使用设计的DNA结构作为测试目标。为此目的，DNA结构被设计成在小于衍射极限的一定间隔处具有结合位点。该结构被标记有几种类型的量子点（QD），以获得它们的空间信息作为光谱编码图像。用点扩散函数多重拟合算法分析所获得的图像，以确定指示结合位点位置的QD位置。实验结果表明，标记的位置可以观察到超过衍射极限的分辨率使用三色荧光图像，获得了共聚焦荧光显微镜。数值模拟表明，标记与八种类型的量子点，使在27.2纳米间距的DNA结构上对齐的位置，以高精度解决。
This paper presents an experimental assessment and analysis of super-resolution microscopy based on multiple-point spread function fitting of spectrally demultiplexed images using a designed DNA structure as a test target. For the purpose, a DNA structure was designed to have binding sites at a certain interval that is smaller than the diffraction limit. The structure was labeled with several types of quantum dots (QDs) to acquire their spatial information as spectrally encoded images. The obtained images are analyzed with a point spread function multifitting algorithm to determine the QD locations that indicate the binding site positions. The experimental results show that the labeled locations can be observed beyond the diffraction-limited resolution using three-colored fluorescence images that were obtained with a confocal fluorescence microscope. Numerical simulations show that labeling with eight types of QDs enables the positions aligned at 27.2-nm pitches on the DNA structure to be resolved with high accuracy.