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
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
The mammalian brain is a highly complex network that consists of millions to billions of densely-interconnected neurons.Precise dissection of neural circuits at the mesoscopic level can provide important structural in...The mammalian brain is a highly complex network that consists of millions to billions of densely-interconnected neurons.Precise dissection of neural circuits at the mesoscopic level can provide important structural information for understanding the brain.Optical approaches can achieve submicron lateral resolution and achieve“optical sectioning”by a variety of means,which has the natural advantage of allowing the observation of neural circuits at the mesoscopic level.Automated whole-brain optical imaging methods based on tissue clearing or histological sectioning surpass the limitation of optical imaging depth in biological tissues and can provide delicate structural information in a large volume of tissues.Combined with various fluorescent labeling techniques,whole-brain optical imaging methods have shown great potential in the brain-wide quantitative profiling of cells,circuits,and blood vessels.In this review,we summarize the principles and implementations of various whole-brain optical imaging methods and provide some concepts regarding their future development.展开更多
The contribution deals with the experimental and numerical investigation of compressible flow through the tip-section turbine blade cascade with the blade 54″ long. Experimental investigations by means of optical(int...The contribution deals with the experimental and numerical investigation of compressible flow through the tip-section turbine blade cascade with the blade 54″ long. Experimental investigations by means of optical(interferometry and schlieren method) and pneumatic measurements provide more information about the behaviour and nature of basic phenomena occurring in the profile cascade flow field. The numerical simulation was carried out by means of the EARSM turbulence model according to Hellsten [5] completed by the bypass transition model with the algebraic equation for the intermittency coefficient proposed by Straka and P?íhoda [6] and implemented into the in-house numerical code. The investigation was focused particularly on the effect of shock waves on the shear layer development including the laminar/turbulent transition. Interactions of shock waves with shear layers on both sides of the blade result usually in the transition in attached and/ or separated flow and so to the considerable impact to the flow structure and energy losses in the blade cascade.展开更多
Wide-field photobleaching imprinting microscopy(PIM)can improve fluorescence image contrast by cleverly exploiting the fluorophores’photobleaching properties.However,as conventional wide-field PIM commonly adopts Gau...Wide-field photobleaching imprinting microscopy(PIM)can improve fluorescence image contrast by cleverly exploiting the fluorophores’photobleaching properties.However,as conventional wide-field PIM commonly adopts Gaussian illumination with a nonuniform lateral fluence distribution,the field-of-view(FOV)and sampling density are largely reduced.In addition,the slow axial fluence gradient of Gaussian illumination limits the signal-to-background ratio(SBR)improvement and optical sectioning capability of PIM.Here,we present flat-field photobleaching imprinting microscopy(ffPIM)with a uniform lateral excitation fluence and sharp axial intensity gradient at the focal plane.ffPIM demonstrates low background,large FOV,and thin optical section.More importantly,compared to either conventional wide-field PIM or light-sheet microscopy,ffPIM shows much better balance for FOV,sampling density,SBR,and optical sectioning capability.The performance of ffPIM is characterized by simulation and resolving multiple cellular structures.Finally,ffPIM can be easily implemented to a standard commercial wide-field microscope and,thereby,allow general laboratories to benefit from this technique.展开更多
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.
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 STI2030-Major Projects(2021ZD0201001 and 2021ZD0201000)the National Natural Science Foundation of China(81827901 and 32192412).
文摘The mammalian brain is a highly complex network that consists of millions to billions of densely-interconnected neurons.Precise dissection of neural circuits at the mesoscopic level can provide important structural information for understanding the brain.Optical approaches can achieve submicron lateral resolution and achieve“optical sectioning”by a variety of means,which has the natural advantage of allowing the observation of neural circuits at the mesoscopic level.Automated whole-brain optical imaging methods based on tissue clearing or histological sectioning surpass the limitation of optical imaging depth in biological tissues and can provide delicate structural information in a large volume of tissues.Combined with various fluorescent labeling techniques,whole-brain optical imaging methods have shown great potential in the brain-wide quantitative profiling of cells,circuits,and blood vessels.In this review,we summarize the principles and implementations of various whole-brain optical imaging methods and provide some concepts regarding their future development.
基金supported by the Technology Agency of the Czech Republic under the grant TA03020277by the Czech Science Foundation under grant P101/12/1271
文摘The contribution deals with the experimental and numerical investigation of compressible flow through the tip-section turbine blade cascade with the blade 54″ long. Experimental investigations by means of optical(interferometry and schlieren method) and pneumatic measurements provide more information about the behaviour and nature of basic phenomena occurring in the profile cascade flow field. The numerical simulation was carried out by means of the EARSM turbulence model according to Hellsten [5] completed by the bypass transition model with the algebraic equation for the intermittency coefficient proposed by Straka and P?íhoda [6] and implemented into the in-house numerical code. The investigation was focused particularly on the effect of shock waves on the shear layer development including the laminar/turbulent transition. Interactions of shock waves with shear layers on both sides of the blade result usually in the transition in attached and/ or separated flow and so to the considerable impact to the flow structure and energy losses in the blade cascade.
基金supported by grants from the National Key R&D Program of China Nos.2021YFF0700304 and 2022YFA3401100 and the National Science Foundation of China 22127804.
文摘Wide-field photobleaching imprinting microscopy(PIM)can improve fluorescence image contrast by cleverly exploiting the fluorophores’photobleaching properties.However,as conventional wide-field PIM commonly adopts Gaussian illumination with a nonuniform lateral fluence distribution,the field-of-view(FOV)and sampling density are largely reduced.In addition,the slow axial fluence gradient of Gaussian illumination limits the signal-to-background ratio(SBR)improvement and optical sectioning capability of PIM.Here,we present flat-field photobleaching imprinting microscopy(ffPIM)with a uniform lateral excitation fluence and sharp axial intensity gradient at the focal plane.ffPIM demonstrates low background,large FOV,and thin optical section.More importantly,compared to either conventional wide-field PIM or light-sheet microscopy,ffPIM shows much better balance for FOV,sampling density,SBR,and optical sectioning capability.The performance of ffPIM is characterized by simulation and resolving multiple cellular structures.Finally,ffPIM can be easily implemented to a standard commercial wide-field microscope and,thereby,allow general laboratories to benefit from this technique.
