表征(材料科学)
反向
点(几何)
显微镜
反问题
点扩散函数
数学
物理
材料科学
数学分析
光学
几何学
作者
Sheng Liu,Jianwei Chen,Jonas Hellgoth,Lucas-Raphael Müller,Boris Ferdman,Dafei Xiao,Keith A. Lidke,Rainer Heintzmann,Yoav Shechtman,Yiming Li,Jonas Ries
标识
DOI:10.1016/j.bpj.2023.11.1806
摘要
The point spread function (PSF) of a microscope describes the image of a point emitter. Knowing the accurate PSF model is essential for various imaging tasks, including single molecule localization, aberration correction and deconvolution. Here we present uiPSF (universal inverse modelling of point spread functions), a toolbox to infer accurate PSF models from microscopy data, using either image stacks of fluorescent beads or directly images of blinking fluorophores, the raw data in single molecule localization microscopy (SMLM). The resulting PSF model enables accurate 3D super-resolution imaging using SMLM. Additionally, uiPSF can be used to characterize and optimize a microscope system where it quantifies the aberrations, including field-dependent aberrations, and the full width half maximum and Strehl ratio of the PSF model. Our modular framework is applicable to a variety of microscope modalities and the PSF model incorporates system or sample specific characteristics, e.g. the bead size, depth dependent aberrations and transformations among channels. We demonstrate its application in single or multiple channels or large field-of-view SMLM systems, 4Pi-SMLM, and lattice light-sheet microscope using either bead data or single molecule blinking data.
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