Recent progress on the reconstruction algorithms of structured illumination microscopy
doi:10.37188/CO.EN.2022-0011
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摘要:
作为现代超分辨成像技术的早期组成部分,结构照明显微镜(SIM)已经发展了近20年。其近期在活细胞中实现了高达60 nm和564 Hz的最佳时空分辨率组合,但也存在一些源于内在重建过程的缺点。本文综述了SIM技术的最新进展,包括超分辨率(SR)重建算法、性能评估及SIM与其他成像技术的集成,以便为生物学家提供实用指导。
Abstract:As an early component of modern Super-Resolution (SR) imaging technology, Structured Illumination Microscopy (SIM) has been developed for nearly twenty years. With up to ~60 nm wavelengths and 564 Hz frame rates, it has recently achieved an optimal combination of spatiotemporal resolution in live cells. Despite these advantages, SIM also suffers disadvantages, some of which originated from the intrinsic reconstruction process. Here we review recent technical advances in SIM, including SR reconstruction, performance evaluation, and its integration with other technologies to provide a practical guide for biologists.
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Figure 1.Schematic diagram of structured illumination microscopy. (a) In sinusoidal illumination microscopy, interference between multiple beams (usually generated by a diffraction grating or spatial light modulator) creates a 2D or 3D striped pattern with spatial frequency
${k}_{{\rm{ex}}}$ illuminating on the sample. This pattern shifts the sample's spatial frequency spectrum$ \stackrel{~}{S}\left(k\right) $ to$\stackrel{~}{S}\left(k+{k}_{{\rm{ex}}}\right)$ and$\stackrel{~}{S}\left(k-{k}_{{\rm{ex}}}\right)$ , translating high-frequency SR information into the diffraction-limited detection passband${OTF}_{{\rm{em}}}\left(k\right)$ with the spatial cutoff frequency${k}_{{\rm{em}}}$ . After computational processing, the sample's highest detectable frequency can be extended to${k}_{{\rm{ex}}}+{k}_{{\rm{em}}}$ . (b) Spot-scanning illumination microscopy where fluorescence is collected by an array detector, and pixels offset by a distance from the excitation spot detect a shifted but higher-resolution, low-signal confocal image. The reconstruction algorithm corrects the shift and restores the signal by reassigning the detected fluorescence toward the illumination axis, with the final resolution${PS F}_{{\rm{sys}}}$ determined by the product of the excitation PSF (${PS F}_{{\rm{ex}}}$ ) and the emission PSF (${PS F}_{{\rm{em}}}$ ). After deconvolution, this process improves resolution similar to that obtained with sinusoidal illumination microscopyFigure 4.(a) Schematic diagram of 3D STED-SIM. (b) The cross-section comparison of lateral PSF (top, left), axial PSF (bottom, left), lateral OTF (top, right), and axial OTF (bottom, right) of the widefield microscopy (red) and 3D STED-SIM (blue). Adapted from Xueet al.[49]
Table 1.Comparison of SIM SR reconstruction algorithm
Principle Effect Code Reference TV-SIM Append TV regularization to reconstruction Suppress reconstruction artifacts Not open-source Chuet al. 2014[14] Hessian-SIM Append Hessian regularization to reconstruction Suppress reconstruction artifacts, avoid over-sharpening boundaries Open-source Huanget al. 2018[15] HiFi-SIM Engineering the effective SIM PSF into an ideal form Suppress reconstruction artifacts, improve axial sectioning Open-source Wenet al. 2021[17] Sparse-SIM Append Sparse and Hessian regularization to reconstruction Increases SIM resolution ~2-fold laterally Open-source Zhaoet al. 2021[26] sCMOS Noise-corrected SIM Introduce sCMOS imaging noise model to reconstruction Suppress sCMOS noise-induced reconstruction artifacts Not open source Zhouet al. 2022[16] Two-step RL deconvolution SIM Introduce two-step RL deconvolution to reconstruction eliminate ad hoc tuneable parameters Not open source Perezet al. 2016[18] Noise-controlled SIM Introduce a physically realistic noise model to reconstruction Suppress reconstruction artifacts, eliminate ad hoc tuneable parameters, maintain resolution and contrast Open-source Smithet al. 2021[19] GAN TIRF-SIM Use GAN for transforming TIRF images into TIRF SIM images Reconstruct rapidly Open-source Wanget al. 2019[20] U-Net SIM Use U-net for producing SIM images Train efficiently and reconstruct with fewer low-intensity input images Open-source Jinet al. 2020[21] 3D RCAN Use 3D RCAN for increasing SIM resolution Increases SIM resolution ~1.9-fold laterally and ~3.6-fold axially Open-source Chenet al. 2021[23] DFCAN/DFGAN Use DFCAN/DFGAN for producing SIM images Reconstruct with low SNR input images Open-source Qiaoet al. 2021[24] Table 2.Summary of SIM performance evaluation algorithms
Function Code Reference FRC/FSC Determine SIM resolution by cross-correlation Open-source Nieuwenhuizenet al. 2013[28] Decorrelation analysis Determine SIM resolution by partial phase correlation Open-source Desclouxet al. 2019[31] NanoJ-SQUIRREL Evaluate SIM artifacts with the resolution scaling function Open-source Culleyet al. 2018[32] SIMcheck Evaluate SIM stripe modulation contrast by computing the standard deviation Open-source Ballet al. 2015[33] -
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