Fluorescence polarization is related to the dipole orientation of chromophores,making fuores-cence polarization microscopy possible to_reveal structures and functions of tagged cellularorganelles and biological macrom...Fluorescence polarization is related to the dipole orientation of chromophores,making fuores-cence polarization microscopy possible to_reveal structures and functions of tagged cellularorganelles and biological macromolecules.Several recent super resolution techniques have beenapplied to fluorescence polarization microscopy,achieving dipole measurement at nanoscale.In this review,we summarize both difraction limited and super resolution fluorescence polari-zation microscopy techniques,as well as their applications in biological imaging.展开更多
A stimulated emission depletion （STED） microscopy scheme using axially symmetric polarized vortex beams is pro- posed based on unique focusing properties of such kinds of beams. The concept of axially symmetric pola...A stimulated emission depletion （STED） microscopy scheme using axially symmetric polarized vortex beams is pro- posed based on unique focusing properties of such kinds of beams. The concept of axially symmetric polarized vortex beams is first introduced, and the basic principle about the scheme is described. Simulation results for several typical beams are then shown, including radially polarized vortex beams, azimuthally polarized vortex beams, and high-order axi- ally symmetric polarized vortex beams. The results indicate that sharper doughnut spots and thus higher resolutions can be achieved, showing more flexibility than previous schemes based on flexible modulation of both phase and polarization for incident beams.展开更多
A super-focusing device composed of a focusing objective and a hyperlens is proposed to focus an incident plane wave into the deep subwavelength dimension. In the device, the objective converts the incident plane wave...A super-focusing device composed of a focusing objective and a hyperlens is proposed to focus an incident plane wave into the deep subwavelength dimension. In the device, the objective converts the incident plane wave into a convergent one. The half cylindrical hyperlens can support high wave vector k modes propagating towards its core. So the convergent wave can be focused into an ultrasmall spot beyond the diffraction limit. The layout is proposed for the super-focusing device and its characteristics are investigated theoretically. Numerical simulations verify that the focused beams are confined in a spot with a diameter of 16.3 nm in the focal plane of the focusing objective with a numerical aperture of 0.6, which corresponds to a super-resolution spot of λ0/23 （λ0 is the wavelength in vacuum）. The simulations confirm the effectiveness of the proposed device.展开更多
Optical microscopy plays an essential role in biological studies due to its capability and compatibility of non-contact,minimally invasive observation and measurement of live specimens.However,the conventional optical...Optical microscopy plays an essential role in biological studies due to its capability and compatibility of non-contact,minimally invasive observation and measurement of live specimens.However,the conventional optical microscopy only has a spatial resolution about200 nm due to the Abbe diffraction limit,and also lacks the ability of three-dimensional imaging.Super-resolution farfield optical microscopy based on special illumination schemes has been dramatically developed over the last decade.Among them,only the structured illumination microscopy(SIM)is of wide-field geometry that enables it easily compatible with the conventional optical microscope.In this article,the principle of SIM was introduced in terms of point spread function(PSF)and optical transform function(OTF)of the optical system.The SIM for super-resolution(SIM-SR)proposed by Gustafsson et al.and the SIM for optical sectioning(SIM-OS)proposed by Neil et al.are the most popular ones in the research community of microscopy.They have the same optical configuration,but with different data postprocessing algorithms.We mathematically described the basic theories for both of the SIMs,respectively,and presented some numerical simulations to show the effects of super-resolution and optical sectioning.Various approaches to generation of structured illumination patterns were reviewed.As an example,a SIM system based on DMDmodulation and LED-illumination was demonstrated.A lateral resolution of 90 nm was achieved with gold nanoparticles.The optical sectioning capability of the microscope was demonstrated with Golgi-stained mouse brain neurons,and the sectioning strength of 930 nm was obtained.展开更多
The resolution of conventional light microscopy is insufficient for subcelluar studies.The invention of various super-resolution imaging techniques breaks the diffraction barrier and pushes the resolution limit toward...The resolution of conventional light microscopy is insufficient for subcelluar studies.The invention of various super-resolution imaging techniques breaks the diffraction barrier and pushes the resolution limit towards the nanometer scale.Here,we focus on a category of super-resolution microscopy that relies on the stochastic activation and precise localization of single molecules.A diversity of fluorescent probes with different characteristics has been developed to achieve super-resolution imaging.In addition,with the implementation of robust localization algorithms,this family of approaches has been expanded to multi-color,three-dimensional and live cell imaging,which provides a promising prospect in biological research.展开更多
Investigating the fine structure of mitochondria and their dynamic interactions with other organelles is crucial for unraveling the mechanisms underlying mitochondrial-related diseases.The development of super-resolut...Investigating the fine structure of mitochondria and their dynamic interactions with other organelles is crucial for unraveling the mechanisms underlying mitochondrial-related diseases.The development of super-resolution techniques has provided powerful visualization tools for mitochondrial research,which is significant for investigating mitochondrial cristae structure,the localization of mitochondrial-related protein complex,and the interactions between mitochondria and other organelles.In this perspective,we introduce several advanced super-resolution techniques and their applications in mitochondrial research,and discuss the potential roles these techniques may play in future studies of mitochondria.展开更多
As a superlens to overcome the well-known diffraction limit,the hyperlens has received much attention due to its super resolving power and magnifying capabilities.In this article,we review the recent developments,incl...As a superlens to overcome the well-known diffraction limit,the hyperlens has received much attention due to its super resolving power and magnifying capabilities.In this article,we review the recent developments,including theoretical and experimental studies on the hyperlens.We also discuss its limitations and potential.展开更多
基金supported by the National Instrument Development Special Program(2013YQ03065102)the Natural Science Foundation of China(614-75010,61428501)Science and Technology Commission of Shanghai Municipality(16DZ-1100300).
文摘Fluorescence polarization is related to the dipole orientation of chromophores,making fuores-cence polarization microscopy possible to_reveal structures and functions of tagged cellularorganelles and biological macromolecules.Several recent super resolution techniques have beenapplied to fluorescence polarization microscopy,achieving dipole measurement at nanoscale.In this review,we summarize both difraction limited and super resolution fluorescence polari-zation microscopy techniques,as well as their applications in biological imaging.
基金Project supported by the National Natural Science Foundation of China(Grant Nos.61108047 and 61475021)the Natural Science Foundation of Beijing,China(Grant No.4152015)+2 种基金the Program for New Century Excellent Talents in Universities of China(Grant No.NCET-13-0667)the Top Young Talents Support Program of BeijingChina(Grant No.CIT&TCD201404113)
文摘A stimulated emission depletion （STED） microscopy scheme using axially symmetric polarized vortex beams is pro- posed based on unique focusing properties of such kinds of beams. The concept of axially symmetric polarized vortex beams is first introduced, and the basic principle about the scheme is described. Simulation results for several typical beams are then shown, including radially polarized vortex beams, azimuthally polarized vortex beams, and high-order axi- ally symmetric polarized vortex beams. The results indicate that sharper doughnut spots and thus higher resolutions can be achieved, showing more flexibility than previous schemes based on flexible modulation of both phase and polarization for incident beams.
基金supported by the National Natural Science Foundation of China(Grant No.10904118)the Natural Science Foundation of Hubei Province,China(Grant No.2009CDB211)the Fundamental Research Funds for the Central Universities of China
文摘A super-focusing device composed of a focusing objective and a hyperlens is proposed to focus an incident plane wave into the deep subwavelength dimension. In the device, the objective converts the incident plane wave into a convergent one. The half cylindrical hyperlens can support high wave vector k modes propagating towards its core. So the convergent wave can be focused into an ultrasmall spot beyond the diffraction limit. The layout is proposed for the super-focusing device and its characteristics are investigated theoretically. Numerical simulations verify that the focused beams are confined in a spot with a diameter of 16.3 nm in the focal plane of the focusing objective with a numerical aperture of 0.6, which corresponds to a super-resolution spot of λ0/23 （λ0 is the wavelength in vacuum）. The simulations confirm the effectiveness of the proposed device.
文摘Optical microscopy plays an essential role in biological studies due to its capability and compatibility of non-contact,minimally invasive observation and measurement of live specimens.However,the conventional optical microscopy only has a spatial resolution about200 nm due to the Abbe diffraction limit,and also lacks the ability of three-dimensional imaging.Super-resolution farfield optical microscopy based on special illumination schemes has been dramatically developed over the last decade.Among them,only the structured illumination microscopy(SIM)is of wide-field geometry that enables it easily compatible with the conventional optical microscope.In this article,the principle of SIM was introduced in terms of point spread function(PSF)and optical transform function(OTF)of the optical system.The SIM for super-resolution(SIM-SR)proposed by Gustafsson et al.and the SIM for optical sectioning(SIM-OS)proposed by Neil et al.are the most popular ones in the research community of microscopy.They have the same optical configuration,but with different data postprocessing algorithms.We mathematically described the basic theories for both of the SIMs,respectively,and presented some numerical simulations to show the effects of super-resolution and optical sectioning.Various approaches to generation of structured illumination patterns were reviewed.As an example,a SIM system based on DMDmodulation and LED-illumination was demonstrated.A lateral resolution of 90 nm was achieved with gold nanoparticles.The optical sectioning capability of the microscope was demonstrated with Golgi-stained mouse brain neurons,and the sectioning strength of 930 nm was obtained.
基金supported by the National Natural Science Foundation of China(31130065,31127002,31100615 and 31100596)the Chinese Academy of Sciences Project(yg2012029)
文摘The resolution of conventional light microscopy is insufficient for subcelluar studies.The invention of various super-resolution imaging techniques breaks the diffraction barrier and pushes the resolution limit towards the nanometer scale.Here,we focus on a category of super-resolution microscopy that relies on the stochastic activation and precise localization of single molecules.A diversity of fluorescent probes with different characteristics has been developed to achieve super-resolution imaging.In addition,with the implementation of robust localization algorithms,this family of approaches has been expanded to multi-color,three-dimensional and live cell imaging,which provides a promising prospect in biological research.
基金supported by the National Science and Technology Major Project Program(2022YFF0712503 and 2021YFA1100201)the National Natural Science Foundation of China(32322049,32350009,32170691 and 92150301)the Beijing Nova Program(20220484033)。
文摘Investigating the fine structure of mitochondria and their dynamic interactions with other organelles is crucial for unraveling the mechanisms underlying mitochondrial-related diseases.The development of super-resolution techniques has provided powerful visualization tools for mitochondrial research,which is significant for investigating mitochondrial cristae structure,the localization of mitochondrial-related protein complex,and the interactions between mitochondria and other organelles.In this perspective,we introduce several advanced super-resolution techniques and their applications in mitochondrial research,and discuss the potential roles these techniques may play in future studies of mitochondria.
文摘超分辨结构光照明显微成像技术(super-resolution structured illumination microscopy,SR-SIM)具有时间分辨率高、光漂白和光毒性低和对荧光探针的要求少等优点,适用于活细胞的长时程超分辨成像.采用二维晶格结构光作为照明光,可以实现更快的成像速度和更低的光毒性,但同时也增加了系统的复杂性.为了解决此问题,本文提出了一种基于数字微镜器件的快速超分辨晶格结构光照明显微成像方法(digital micromirror device-based lattice SIM,DMD-Lattice-SIM),通过同步分时触发DMD和sCMOS相机的方式实现二维正交晶格结构光的产生,且只需要采集5幅相移原始图像即可重构出超分辨图像,相比于传统SR-SIM需要9幅相移原始图像的方法,图像采集时间减少了约44.4%.同时,在基于空域和频域联合的SIM重构算法(joint space and frequency reconstruction method-SIM,JSFR-SIM)的基础上,本文还发展了用于Lattice-SIM的JSFR超分辨图像重构方法(Lattice-JSFR-SIM),先在频域对原始图像进行预滤波处理;然后,在空域对滤波后的图像进行超分辨重构处理.与传统频域图像重构处理对比,该方法在512×512像素数的成像视场下重构时间减少了约55.6%,对于实现活细胞实时超分辨成像具有重要意义和应用价值.
文摘As a superlens to overcome the well-known diffraction limit,the hyperlens has received much attention due to its super resolving power and magnifying capabilities.In this article,we review the recent developments,including theoretical and experimental studies on the hyperlens.We also discuss its limitations and potential.
