We report a comprehensive numerical study of super resolution (SR) structured illumination microscopy (SIM) utilizing the classic Heintzmann-Cremer SIM process and algorithm. In particular, we investigated the impact ...We report a comprehensive numerical study of super resolution (SR) structured illumination microscopy (SIM) utilizing the classic Heintzmann-Cremer SIM process and algorithm. In particular, we investigated the impact of the diffraction limit of the underlying imaging system on the optimal SIM grating frequency that can be used to obtain the highest SR enhancement with non-continuous spatial frequency support. Besides confirming the previous theoretical and experimental work that SR-SIM can achieve an enhancement close to 3 times the diffraction limit with grating pattern illuminations, we also observe and report a series of more subtle effects of SR-SIM with non-continuous spatial frequency support. Our simulations show that when the SIM grating frequency exceeds twice that of the diffraction limit, the higher SIM grating frequency can help achieve a higher SR enhancement for the underlying imaging systems whose diffraction limit is low, though this enhancement is obtained at the cost of losing resolution at some lower resolution targets. Our simulations also show that, for underlying imaging systems with high diffraction limits, however, SR-SIM grating frequencies above twice the diffraction limits tend to bring no significant extra enhancement. Furthermore, we observed that there exists a limit grating frequency above which the SR enhancement effect is lost, and the reconstructed images essentially have the same resolution as the one obtained directly from the underlying imaging system without using the SIM process.展开更多
Structured illumination microscopy has been a useful method for achieving lateral super-resolution,but it typically requires at least three precise phase shifts per orientation.In this paper,we propose a super-resolut...Structured illumination microscopy has been a useful method for achieving lateral super-resolution,but it typically requires at least three precise phase shifts per orientation.In this paper,we propose a super-resolution method that utilizes structured illumination without phase shift.The reconstruction process requires only a conventionally illuminated image and an image with structured illumination.This method achieves the same effect as the traditional phase shift method,and more than doubles the resolution by synthesizing a few reconstructions at different illumination frequencies.We verify the resolution improvement process using a combination of theoretical derivations and diagrams,and demonstrate its effectiveness with numerical simulations.展开更多
A maximum a posteriori( MAP) algorithm is proposed to improve the accuracy of super resolution( SR) reconstruction in traditional methods. The algorithm applies both joints image registration and SR reconstruction...A maximum a posteriori( MAP) algorithm is proposed to improve the accuracy of super resolution( SR) reconstruction in traditional methods. The algorithm applies both joints image registration and SR reconstruction in the framework,but separates them in the process of iteratiion. Firstly,we estimate the shifting parameters through two lowresolution( LR) images and use the parameters to reconstruct initial HR images. Then,we update the shifting parameters using HR images. The aforementioned steps are repeated until the ideal HR images are obtained. The metrics such as PSNR and SSIM are used to fully evaluate the quality of the reconstructed image. Experimental results indicate that the proposed method can enhance image resolution efficiently.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
文摘We report a comprehensive numerical study of super resolution (SR) structured illumination microscopy (SIM) utilizing the classic Heintzmann-Cremer SIM process and algorithm. In particular, we investigated the impact of the diffraction limit of the underlying imaging system on the optimal SIM grating frequency that can be used to obtain the highest SR enhancement with non-continuous spatial frequency support. Besides confirming the previous theoretical and experimental work that SR-SIM can achieve an enhancement close to 3 times the diffraction limit with grating pattern illuminations, we also observe and report a series of more subtle effects of SR-SIM with non-continuous spatial frequency support. Our simulations show that when the SIM grating frequency exceeds twice that of the diffraction limit, the higher SIM grating frequency can help achieve a higher SR enhancement for the underlying imaging systems whose diffraction limit is low, though this enhancement is obtained at the cost of losing resolution at some lower resolution targets. Our simulations also show that, for underlying imaging systems with high diffraction limits, however, SR-SIM grating frequencies above twice the diffraction limits tend to bring no significant extra enhancement. Furthermore, we observed that there exists a limit grating frequency above which the SR enhancement effect is lost, and the reconstructed images essentially have the same resolution as the one obtained directly from the underlying imaging system without using the SIM process.
基金supported by the National Natural Science Foundation of China(Grant No.51775381)National Key Scientific Instrument and Equipment Development Project(Grant No.2017YFF0107001)
文摘Structured illumination microscopy has been a useful method for achieving lateral super-resolution,but it typically requires at least three precise phase shifts per orientation.In this paper,we propose a super-resolution method that utilizes structured illumination without phase shift.The reconstruction process requires only a conventionally illuminated image and an image with structured illumination.This method achieves the same effect as the traditional phase shift method,and more than doubles the resolution by synthesizing a few reconstructions at different illumination frequencies.We verify the resolution improvement process using a combination of theoretical derivations and diagrams,and demonstrate its effectiveness with numerical simulations.
基金Supported by the National Natural Science Foundation of China(61405191)
文摘A maximum a posteriori( MAP) algorithm is proposed to improve the accuracy of super resolution( SR) reconstruction in traditional methods. The algorithm applies both joints image registration and SR reconstruction in the framework,but separates them in the process of iteratiion. Firstly,we estimate the shifting parameters through two lowresolution( LR) images and use the parameters to reconstruct initial HR images. Then,we update the shifting parameters using HR images. The aforementioned steps are repeated until the ideal HR images are obtained. The metrics such as PSNR and SSIM are used to fully evaluate the quality of the reconstructed image. Experimental results indicate that the proposed method can enhance image resolution efficiently.
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.
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.