The originally published version of this paper regrettably contained some typos.First,“structure illumination microscopy”should have been written as“structured illumination microscopy”throughout the text,including...The originally published version of this paper regrettably contained some typos.First,“structure illumination microscopy”should have been written as“structured illumination microscopy”throughout the text,including in the article title,graphical abstract,the summary,and the main text.Second,in Figure 1A,“iFFT”should be written as“FFT.”Third,in Video S2,the labels“FO”and“Open”were placed incorrectly;FO is the high-quality reconstruction result,while Open contains reconstruction artifact.展开更多
Three-dimensional structured illumination microscopy(3DSIM)is a popular method for observing subcellular/cellular structures or animal/plant tissues with gentle phototoxicity and 3D super-resolution.However,its time-c...Three-dimensional structured illumination microscopy(3DSIM)is a popular method for observing subcellular/cellular structures or animal/plant tissues with gentle phototoxicity and 3D super-resolution.However,its time-consuming reconstruction process poses challenges for high-throughput imaging and real-time observation.Moreover,traditional 3DSIM typically requires more than six z layers for successful reconstruction and is susceptible to defocused backgrounds.This poses a great gap between single-layer 2DSIM and 6-layer 3DSIM,and limits the observation of thicker samples.To address these limitations,we developed FO-3DSIM,a novel method that integrates spatial-domain reconstruction with optical-sectioning SIM.FO-3DSIM enhances reconstruction speed by up to 855.7 times with superior performance with limited z layers and under high defocused backgrounds.It retains the high-fidelity,low-photon reconstruction capabilities of our previously proposed Open-3DSIM.Utilizing fast reconstruction and optical sectioning,we achieved large field-of-view(FOV)3D super-resolution imaging of mouse kidney actin,covering a region of 0.453 mm×0.453 mm×2.75μm within 23 min of acquisition and 13 min of reconstruction.Near real-time performance was demonstrated in live actin imaging with FO-3DSIM.Our approach reduces photodamage through limited z layer reconstruction,allowing the observation of ER tubes with just three layers.We anticipate that FO-3DSIM will pave the way for near real-time,large FOV 6D imaging,encompassing xyz super-resolution,multi-color,long-term,and polarization imaging with less photodamage,removed defocused backgrounds,and reduced reconstruction time.展开更多
Three-dimensional structured illumination microscopy(3DSIM)is an essential super-resolution imaging technique for visualizing volumetric subcellular structures at the nanoscale,capable of doubling both lateral and axi...Three-dimensional structured illumination microscopy(3DSIM)is an essential super-resolution imaging technique for visualizing volumetric subcellular structures at the nanoscale,capable of doubling both lateral and axial resolution beyond the diffraction limit.However,high-quality 3DSIM reconstruction is often hindered by uncertainties in experimental parameters,such as optical aberrations and fluorescence density heterogeneity.Here,we present PCA-3DSIM,a novel 3DSIM reconstruction framework that extends principal component analysis(PCA)from two-dimensional(2D)to three-dimensional(3D)super-resolution microscopy.To further compensate spatial nonuniformities of illumination parameters,PCA-3DSIM can be implemented in an adaptive tiled-block manner.By segmenting raw volumetric data into localized subsets,PCA-3DSIM enables accurate parameter estimation and effective interference rejection for high-fidelity,artifact-free 3D super-resolution reconstruction,with the inherent efficiency of PCA supporting the tiled reconstruction with limited computational burden.Experimental results demonstrate that PCA-3DSIM provides reliable reconstruction performance and improved robustness across diverse imaging scenarios,from custom-built platforms to commercial systems.These results establish PCA-3DSIM as a flexible and practical tool for super-resolved volumetric imaging of subcellular structures,with broad potential applications in biomedical research.展开更多
Dear Editor,The mammalian brain exhibits cross-scale complexity in neuronal morphology and connectivity,the study of which demands high-resolution morphological reconstruction of individual neurons across the entire b...Dear Editor,The mammalian brain exhibits cross-scale complexity in neuronal morphology and connectivity,the study of which demands high-resolution morphological reconstruction of individual neurons across the entire brain[1-4].Current commonly used approaches for such mesoscale brain mapping include two main types of three-dimensional fluorescence microscopy:the block-face methods,and the lightsheet-based methods[5,6].In general,the high imaging speed and light efficiency of light-sheet microscopy make it a suitable tool for high-throughput volumetric imaging,especially when combined with tissue-clearing techniques.However,large brain samples pose major challenges to this approach.展开更多
文摘The originally published version of this paper regrettably contained some typos.First,“structure illumination microscopy”should have been written as“structured illumination microscopy”throughout the text,including in the article title,graphical abstract,the summary,and the main text.Second,in Figure 1A,“iFFT”should be written as“FFT.”Third,in Video S2,the labels“FO”and“Open”were placed incorrectly;FO is the high-quality reconstruction result,while Open contains reconstruction artifact.
基金supported by the National Key R&D Program of China(2022YFC3401100)the National Natural Science Foundation of China(624B2009,62405010,62335008,62025501,92150301,and 62411540238).
文摘Three-dimensional structured illumination microscopy(3DSIM)is a popular method for observing subcellular/cellular structures or animal/plant tissues with gentle phototoxicity and 3D super-resolution.However,its time-consuming reconstruction process poses challenges for high-throughput imaging and real-time observation.Moreover,traditional 3DSIM typically requires more than six z layers for successful reconstruction and is susceptible to defocused backgrounds.This poses a great gap between single-layer 2DSIM and 6-layer 3DSIM,and limits the observation of thicker samples.To address these limitations,we developed FO-3DSIM,a novel method that integrates spatial-domain reconstruction with optical-sectioning SIM.FO-3DSIM enhances reconstruction speed by up to 855.7 times with superior performance with limited z layers and under high defocused backgrounds.It retains the high-fidelity,low-photon reconstruction capabilities of our previously proposed Open-3DSIM.Utilizing fast reconstruction and optical sectioning,we achieved large field-of-view(FOV)3D super-resolution imaging of mouse kidney actin,covering a region of 0.453 mm×0.453 mm×2.75μm within 23 min of acquisition and 13 min of reconstruction.Near real-time performance was demonstrated in live actin imaging with FO-3DSIM.Our approach reduces photodamage through limited z layer reconstruction,allowing the observation of ER tubes with just three layers.We anticipate that FO-3DSIM will pave the way for near real-time,large FOV 6D imaging,encompassing xyz super-resolution,multi-color,long-term,and polarization imaging with less photodamage,removed defocused backgrounds,and reduced reconstruction time.
基金supported by National Natural Science Foundation of China(62405136,62275125,62275121,12204239,62175109)China Postdoctoral Science Foundation(BX20240486,2024M754141)+3 种基金Youth Foundation of Jiangsu Province(BK20241466,BK20220946)Jiangsu Funding Program for Excellent Postdoctoral Talent(2024ZB671)Fundamental Research Funds for the Central Universities(30925010309,30922010313,2023102001)Open Research Fund of Jiangsu Key Laboratory of Spectral Imaging&Intelligent Sense(JSGPCXZNGZ202402,JSGP202201).
文摘Three-dimensional structured illumination microscopy(3DSIM)is an essential super-resolution imaging technique for visualizing volumetric subcellular structures at the nanoscale,capable of doubling both lateral and axial resolution beyond the diffraction limit.However,high-quality 3DSIM reconstruction is often hindered by uncertainties in experimental parameters,such as optical aberrations and fluorescence density heterogeneity.Here,we present PCA-3DSIM,a novel 3DSIM reconstruction framework that extends principal component analysis(PCA)from two-dimensional(2D)to three-dimensional(3D)super-resolution microscopy.To further compensate spatial nonuniformities of illumination parameters,PCA-3DSIM can be implemented in an adaptive tiled-block manner.By segmenting raw volumetric data into localized subsets,PCA-3DSIM enables accurate parameter estimation and effective interference rejection for high-fidelity,artifact-free 3D super-resolution reconstruction,with the inherent efficiency of PCA supporting the tiled reconstruction with limited computational burden.Experimental results demonstrate that PCA-3DSIM provides reliable reconstruction performance and improved robustness across diverse imaging scenarios,from custom-built platforms to commercial systems.These results establish PCA-3DSIM as a flexible and practical tool for super-resolved volumetric imaging of subcellular structures,with broad potential applications in biomedical research.
基金supported by the STI 2030-Major Project(2021ZD0204400,2022ZD0205203,2021ZD0200104,2022ZD0211900)the Shenzhen Science and Technology Program(RCYX20210706092100003,RCBS20221008093311027)+3 种基金the Shenzhen Medical Research Funds(A2303005)the Youth Innovation Promotion Association CAS(2022367)the National Natural Science Foundation of China(32100896)NSFC-Guangdong Joint Fund(U20A6005).
文摘Dear Editor,The mammalian brain exhibits cross-scale complexity in neuronal morphology and connectivity,the study of which demands high-resolution morphological reconstruction of individual neurons across the entire brain[1-4].Current commonly used approaches for such mesoscale brain mapping include two main types of three-dimensional fluorescence microscopy:the block-face methods,and the lightsheet-based methods[5,6].In general,the high imaging speed and light efficiency of light-sheet microscopy make it a suitable tool for high-throughput volumetric imaging,especially when combined with tissue-clearing techniques.However,large brain samples pose major challenges to this approach.
