Lateral Resolution Enhancement Research Articles

A Structured Illumination Microscope with the Combination of Optical Sectioning and Lateral Resolution Enhancement for Precise 3D Industrial Metrology

A Structured Illumination Microscope with the Combination of Optical Sectioning and Lateral Resolution Enhancement for Precise 3D Industrial Metrology

A Millimeter-Wave Vortex Phase Imaging Method Developed for Lateral Resolution Enhancement

A Millimeter-Wave Vortex Phase Imaging Method Developed for Lateral Resolution Enhancement

Front Cover

A method based on deep learning was proposed to enhance the defocused resolution and signal‐to‐noise ratio of optical‐resolution photoacoustic microscopy. A fully dense U‐Net was trained with randomly distributed sources of different shapes to improve the quality of photoacoustic images. The results show that the PSNR of defocused signal was enhanced by more than 1.2 times. An over 2.6‐fold enhancement in lateral resolution and an over 3.4‐fold enhancement in axial resolution of defocused regions were achieved.For further details please visit the article by Rui Wang, Zhipeng Zhang, Ruiyi Chen, Xiaohai Yu, Hongyu Zhang, Gang Hu, Qiegen Liu, Xianlin Song (e202300149). image

Noise-insensitive defocused signal and resolution enhancement for optical-resolution photoacoustic microscopy via deep learning.

Optical-resolution photoacoustic microscopy suffers from narrow depth of field and a significant deterioration in defocused signal intensity and spatial resolution. Here, a method based on deep learning was proposed to enhance the defocused resolution and signal-to-noise ratio. A virtual optical-resolution photoacoustic microscopy based on k-wave was used to obtain the datasets of deep learning with different noise levels. A fully dense U-Net was trained with randomly distributed sources to improve the quality of photoacoustic images. The results show that the PSNR of defocused signal was enhanced by more than 1.2 times. An over 2.6-fold enhancement in lateral resolution and an over 3.4-fold enhancement in axial resolution of defocused regions were achieved. The large volumetric and high-resolution imaging of blood vessels further verified that the proposed method can effectively overcome the deterioration of the signal and the spatial resolution due to the narrow depth of field of optical-resolution photoacoustic microscopy.

Restoring the imaging quality of circular transducer array-based PACT using synthetic aperture focusing technique integrated with 2nd-derivative-based back projection scheme

Circular-array-based photoacoustic computed tomography (CA-PACT) is a promising imaging tool owing to its broad acoustic detection coverage and fidelity. However, CA-PACT suffers from poor image quality outside the focal zone along both elevational and lateral dimensions. To address this challenge, we proposed a novel reconstruction strategy by integrating the synthetic aperture focusing technique (SAFT) with the 2nd derivative-based back projection (2nd D-BP) algorithm to restore the image quality outside the focal zone along both the elevational and lateral axes. The proposed solution is a two-phase reconstruction scheme. In the first phase, with the assistance of an acoustic lens, we designed a circular array-based SAFT algorithm to restore the resolution and SNR along the elevational axis. The acoustic lens pushes the boundary of the upper limit of the SAFT scheme to achieve enhanced elevational resolution. In the second phase, we proposed a 2nd D-BP scheme to improve the lateral resolution and suppress noises in 3D imaging results. The 2nd D-BP strategy enhances the image quality along the lateral dimension by up-converting the high spatial frequencies of the object’s absorption pattern. We validated the effectiveness of the proposed strategy using both phantoms and in vivo human experiments. The experimental results demonstrated superior image quality (7-fold enhancement in elevational resolution, 3-fold enhancement in lateral resolution, and an 11-dB increase in SNR). This strategy provides a new paradigm in the PACT system as it significantly enhances the spatial resolution and imaging contrast in both the elevational and lateral dimensions while maintaining a large focal zone.

Hyperbolic metamaterial structures based on graphene for THz super-resolution imaging applications

The diffraction limit of light due to the loss of evanescent waves that carry high spatial frequency information in the far field restricts the practical applications of terahertz imaging technology. In order to break the diffraction limit, we investigate the super-resolution capability of different types of multilayered graphene-dielectric hyperbolic metamaterials. A super-resolution of λ/10 is achieved for both the cylindrical and planar structures. A prominent advantage of graphene-dielectric hyperbolic structures is the dynamic tunability of the dispersion and super-resolution performance by adjusting the chemical potential of graphene through conveniently changing the gate voltage without modifying the geometry of the hyperbolic structures. Furthermore, we have investigated the influence of bilayer thickness variations on the super-resolution performance. Finally, we apply the planar hyperbolic structures for the super-resolution imaging and a roughly five-fold lateral resolution enhancement is realized in our approach. Due to many prominent advantages including super-resolution over broad spectral range, dynamic tunability, good stability and robustness, we believe this work could contribute to the improvement of the resolutions of terahertz imaging systems and the development of hyperbolic metamaterial modulation devices in the terahertz band.

Super-resolution imaging based on radially polarized beam induced superoscillation using an all-dielectric metasurface.

Superoscillation is a kind of phenomenon which can generate oscillation faster than the fastest component of a band-limited function. For optics, superoscillation is generated by coherence of low spatial frequency waves. It can bring a localized region named "hot spot", which has a smaller size than the diffraction-limit, and this character has potential applicaions in super-resolution imaging. Using a high-order radially polarized Laguerre-Gaussian beam tightly focused by high-NA objective lens, we can easily obtain and control the superoscillation hot spot. Using a metasurface, which has compact volume and sub-wavelength pixel size, we can generate the high-order radially polarized Laguerre-Gaussian beam more simply than conventional methods like using a liquid crystal mode converter. We first analyze the properties of unit cells of the metasurface and simulate the performance of the metasurface. Then we analyze the property of the tightly focused high-order radially polarized Laguerre-Gaussian beam and design a super-resolution imaging system using our designed metasurface. Therefore, the 2-fold lateral resolution enhancement is realized in our approach. This method can be used to improve lateral resolution in conventional confocal imaging systems.

High-resolution seismic velocity analysis by sign-based weighted semblance

Building an accurate velocity model plays a vital role in routine seismic imaging workflows. Normal-moveout (NMO)-based seismic velocity analysis is a popular method for making the velocity models. However, traditional velocity analysis methodologies are not generally capable of handling amplitude variations across moveout curves, specifically polarity reversals caused by amplitude-variation-with-offset (AVO) anomalies. I have developed an NMO-based velocity analysis approach that circumvents this shortcoming by modifying the conventional semblance (CS) function to include polarity and amplitude correction terms computed using correlation coefficients of seismic traces in the velocity analysis scanning window with a reference trace. Thus, the proposed method workflow is suitable for any class of AVO effects. The approach is evaluated with four synthetic data examples of different conditions and field data consisting of a common-midpoint gather. Lateral resolution enhancement using the proposed method workflow is evaluated by comparison between the results from the workflow and the results obtained by the application of CS and three semblance-based velocity analysis algorithms developed to circumvent the challenges associated with amplitude variations across moveout curves, caused by seismic attenuation and class II AVO anomalies. According to the obtained results, the proposed method is superior to all of the presented workflows in handling such anomalies.

Enhancement of second harmonic microscopy images in collagen-based thick samples using radially polarized laser beams

The visualization of collagen-based tissues imaged with second harmonic generation (SHG) microscopy strongly depends on the polarization state of the scanning laser beam. This has been often explored using spatially homogeneous states (mainly linear or circular). However, vector beams (i.e. laser beams with spatially non-uniform polarization states) have become a powerful tool in different research field and technological applications. In particular, radially polarized vector beams are known to provide a tight focal spot and lateral resolution enhancement. Here we propose the use of radially polarized beams to improve the quality of SHG microscopy images. The performance of this particular type of vector beam is compared to that obtained with a circularly polarized light beam for different non-stained specimens and depth locations. Results show that independently of the sample’s thickness, the efficiency of SHG processes is increased when using the radially polarized beam, also leading to a higher image quality. Our results open the route for using vector beams to get a better visualization of biological details within the imaged areas, especially at deeper locations.

Three-dimensional residual channel attention networks denoise and sharpen fluorescence microscopy image volumes.

We demonstrate residual channel attention networks (RCAN) for the restoration and enhancement of volumetric time-lapse (four-dimensional) fluorescence microscopy data. First we modify RCAN to handle image volumes, showing that our network enables denoising competitive with three other state-of-the-art neural networks. We use RCAN to restore noisy four-dimensional super-resolution data, enabling image capture of over tens of thousands of images (thousands of volumes) without apparent photobleaching. Second, using simulations we show that RCAN enables resolution enhancement equivalent to, or better than, other networks. Third, we exploit RCAN for denoising and resolution improvement in confocal microscopy, enabling ~2.5-fold lateral resolution enhancement using stimulated emission depletion microscopy ground truth. Fourth, we develop methods to improve spatial resolution in structured illumination microscopy using expansion microscopy data as ground truth, achieving improvements of ~1.9-fold laterally and ~3.6-fold axially. Finally, we characterize the limits of denoising and resolution enhancement, suggesting practical benchmarks for evaluation and further enhancement of network performance.

High-resolution 3D light-field imaging.

.Significance: High-speed 3D imaging methods have been playing crucial roles in many biological discoveries.Aim: We present a hybrid light-field imaging system and image processing algorithm that can visualize high-speed biological events.Approach: The hybrid light-field imaging system uses the selective plane optical illumination, which simultaneously records a high-resolution 2D image and a low-resolution 4D light-field image. The high-resolution 4D light-field image is obtained by applying the hybrid algorithm derived from the deconvolution and phase retrieval methods.Results: High-resolution 3D imaging at a speed of 100-s volumes per second over an imaging field of in the , , and axis, respectively, is achieved with a 2.5 times enhancement in lateral resolution over the entire imaging field compared with standard light-field systems. In comparison to the deconvolution algorithm, the hybrid algorithm addresses the artifact issue at the focal plane and reduces the computation time by a factor of 4.Conclusions: The new hybrid light-field imaging method realizes high-resolution and ultrafast 3D imaging with a compact setup and simple algorithm, which may help discover important applications in biophotonics to visualize high-speed biological events.

SOFISM: Super-resolution optical fluctuation image scanning microscopy

Super-resolution optical microscopy is a rapidly evolving scientific field dedicated to imaging sub-wavelength-sized objects, leaving its mark in multiple branches of biology and technology. While several super-resolution optical microscopy methods have become a common tool in life science imaging, new methods, supported by cutting-edge technology, continue to emerge. One rather recent addition to the super-resolution toolbox, image scanning microscopy (ISM), achieves up to twofold lateral resolution enhancement in a robust and straightforward manner. To further enhance ISM’s resolution in all three dimensions, we present and experimentally demonstrate here super-resolution optical fluctuation ISM (SOFISM). Measuring the fluorescence fluctuation contrast in an ISM architecture, we obtain images with a × 2.5 lateral resolution beyond the diffraction limit along with an enhanced axial resolution for a fixed cell sample labeled with commercially available quantum dots. The inherent temporal averaging of the ISM technique enables image acquisition of the fluctuation correlation contrast within millisecond-scale pixel dwell times. SOFISM can therefore offer a robust path to achieve high-resolution images within a slightly modified confocal microscope, using standard fluorescent labels and reasonable acquisition times.

Resolution-enhanced imaging using interferenceless coded aperture correlation holography with sparse point response

Interferenceless coded aperture correlation holography (I-COACH) is a non-scanning, motionless, incoherent digital holography technique. In this study we use a special type of I-COACH in which its point spread hologram (PSH) is ensemble of sparse dots. With this PSH an imaging resolution beyond the classic diffraction limit is demonstrated. This resolution improvement is achieved due to the position of the coded aperture between the object and the lens-based imaging system. The coded aperture scatters part of the light, that otherwise is blocked by the system aperture, into the optical system, and by doing that, extends the effective numerical aperture of the system. The use of sparse PSH increases the signal-to-noise ratio of the entire imaging system. A lateral resolution enhancement by a factor of about 1.6 was noted in the case of I-COACH compared to direct imaging.

Demonstration of lateral resolution enhancement by focusing amplitude modulated radially polarized light in a confocal imaging system

Lateral resolution enhancement is demonstrated in a confocal imaging system with amplitude-modulated radially polarized (RP) light at the wavelength Annular pupil fields and optimized amplitude distribution functions can be realized with a spatial light modulator. By comparing images obtained with full and amplitude modulated apertures of RP illuminations using a high numerical aperture (NA = 0.9), spatial resolution of has been achieved experimentally. This result agrees very well with theoretical simulation results and will be helpful in improving performance of the non-fluorescent imaging systems.

Enhancement of Acoustic Microscopy Lateral Resolution: A Comparison Between Deep Learning and Two Deconvolution Methods.

Scanning acoustic microscopy (SAM) provides high-resolution images of biological tissues. Since higher transducer frequencies limit penetration depth, image resolution enhancement techniques could help in maintaining sufficient lateral resolution without sacrificing penetration depth. Compared with existing SAM research, this work introduces two novelties. First, deep learning (DL) is used to improve lateral resolution of 180-MHz SAM images, comparing it with two deconvolution-based approaches. Second, 316-MHz images are used as ground truth in order to quantitatively evaluate image resolution enhancement. The samples used were mouse and rat brain sections. The results demonstrate that DL can closely approximate ground truth (NRMSE = 0.056 and PSNR = 28.4 dB) even with a relatively limited training set (four images, each smaller than 1 mm ×1 mm). This study suggests the high potential of using DL as a single image superresolution method in SAM.

Computational Image Enhancement for Frequency Modulated Continuous Wave (FMCW) THz Image

In this paper, a novel method to enhance Frequency Modulated Continuous Wave (FMCW) THz imaging resolution beyond its diffraction limit is proposed. Our method comprises two stages. Firstly, we reconstruct the signal in depth-direction using a sinc-envelope, yielding a significant improvement in depth estimation and signal parameter extraction. The resulting high-precision depth estimate is used to deduce an accurate reflection intensity THz image. This image is fed in the second stage of our method to a 2D blind deconvolution procedure, adopted to enhance the lateral THz image resolution beyond the diffraction limit. Experimental data acquired with a FMCW system operating at 577 GHz with a bandwidth of 126 GHz shows that the proposed method enhances the lateral resolution by a factor of 2.29 to 346.2 μm with respect to the diffraction limit. The depth accuracy is 91 μm. Interestingly, the lateral resolution enhancement achieved with this blind deconvolution concept leads to better results in comparison with conventional gaussian deconvolution. Experimental data on a PCB resolution target is presented, in order to quantify the resolution enhancement and to compare the performance with established image enhancement approaches. The presented technique allows exposure of the interwoven fiber reinforced embedded structures of the PCB test sample.

Microsphere-assisted self-referencing digital holographic microscopy in transmission mode

In this paper, we introduce a self-referencing microsphere-assisted transmission digital holographic microscopy (DHM) system in real-image operating mode. Self-referencing DHM arrangements provide a compact geometry that are temporally stable against environmental vibrations and suitable for the measurement of dynamic specimens such as cells. These advantages may be more pronounced for microsphere-assisted DHM systems. In the real-image mode, unlike the virtual-image mode, the working distance is increased. This, in turn, provides flexibility for insertion of additional elements for enhancement of the image quality, or for other required tasks. The lateral resolution enhancement is similar to the virtual image arrangement and the axial resolution is decoupled from the lateral one. The methodology is discussed theoretically and validated experimentally by conducting DHM experiments on standard micro-objects and aggregation of micro-particles. Our results show that, by the assistance of a rather big size 550 μm silica microsphere, a 10× microscope objective can resolve the 3D structure of a compact disk. The arrangement may be useful toward having a compact and inexpensive bench-top high-resolution three-dimensional imaging apparatus.

Resolution enhancement in nonlinear interferenceless COACH with point response of subdiffraction limit patterns.

Interferenceless coded aperture correlation holography (I-COACH) is a non-scanning, motionless, incoherent digital holography technique for 3D imaging. The lateral and axial resolutions of I-COACH are equivalent to those of conventional direct imaging with the same numerical aperture. The main component of I-COACH is a coded phase mask (CPM) used as the system aperture. In this study, the CPM has been engineered using a modified Gerchberg-Saxton algorithm to generate a random distribution of subdiffraction spot arrays on the digital camera as a system response to a point source illumination. A library of point object holograms is created to calibrate the system for imaging different lateral sections of a 3D object. An object is placed within the calibrated 3D space and an object hologram is recorded with the same CPM. The various planes of the object are reconstructed by a non-linear cross-correlation between the object hologram and the point object hologram library. A lateral resolution enhancement of about 25% was noted in the case of I-COACH compared to direct imaging.

Supercritical focusing coherent anti-Stokes Raman scattering microscopy for high-resolution vibrational imaging.

We report the development of, to the best of our knowledge, a novel supercritical focusing coherent anti-Stokes Raman scattering (SCF-CARS) microscopy for high-resolution vibrational imaging. Two optimized phase patterns with a combination of concentric rings with an alternative 0 and π phase are generated by using a spatial light modulator and applied to the pump beam for minimizing its focal spot size. One of the phase patterns is for both the lateral and axial resolution enhancement, and the other can further improve the lateral resolution, but it sacrifices the axial resolution to some extent. We demonstrate this high-resolution SCF-CARS microscopy technique by imaging the polymethyl methacrylate (PMMA) nano-cylinder on a microscope slide and glass-air interface, as well as biomedical samples, for example, tooth.

Spinning-disc confocal microscopy in the second near-infrared window (NIR-II)

Fluorescence microscopy in the second near-infrared optical window (NIR-II, 1000–1350 nm) has become a technique of choice for non-invasive in vivo imaging. The deep penetration of NIR light in living tissue, as well as negligible tissue autofluorescence within this optical range, offers increased resolution and contrast with even greater penetration depths. Here, we present a custom-built spinning-disc confocal laser microscope (SDCLM) that is specific to imaging in the NIR-II. The SDCLM achieves a lateral resolution of 0.5 ± 0.1 µm and an axial resolution of 0.6 ± 0.1 µm, showing a ~17% and ~45% enhancement in lateral and axial resolution, respectively, compared to the corresponding wide-field configuration. We furthermore showcase several applications that demonstrate the use of the SDCLM for in situ, spatiotemporal tracking of NIR particles and bioanalytes within both synthetic and biological systems.