Resolution Test Target Research Articles

Design and validation of a high-speed hyperspectral laparoscopic imaging system.

Minimally invasive surgery (MIS) has shown vast improvement over open surgery by reducing post-operative stays, intraoperative blood loss, and infection rates. However, in spite of these improvements, there are still prevalent issues surrounding MIS that may be addressed through hyperspectral imaging (HSI). We present a laparoscopic HSI system to further advance the field of MIS. We present an imaging system that integrates high-speed HSI technology with a clinical laparoscopic setup and validate the system's accuracy and functionality. Different configurations that cover the visible (VIS) to near-infrared (NIR) range of electromagnetism are assessed by gauging the spectral fidelity and spatial resolution of each hyperspectral camera. Standard Spectralon reflectance tiles were used to provide ground truth spectral footprints to compare with those acquired by our system using the root mean squared error (RMSE). Demosaicing techniques were investigated and used to measure and improve spatial resolution, which was assessed with a USAF resolution test target. A perception-based image quality evaluator was used to assess the demosaicing techniques we developed. Two configurations of the system were developed for evaluation. The functionality of the system was investigated in a phantom study and by imaging ex vivo tissues. Multiple configurations of our system were tested, each covering different spectral ranges, including VIS (460 to 600nm), red/NIR (RNIR) (610 to 850nm), and NIR (665 to 950nm). Each configuration is capable of achieving real-time imaging speeds of up to 20 frames per second. RMSE values of , , and 3.47% were achieved for the VIS, RNIR, and NIR systems, respectively. We obtained sub-millimeter resolution using our demosaicing techniques. We developed and validated a high-speed hyperspectral laparoscopic imaging system. The HSI system can be used as an intraoperative imaging tool for tissue classification during laparoscopic surgery.

Super-Resolution Image Reconstruction of Wavefront Coding Imaging System Based on Deep Learning Network

Wavefront Coding (WFC) is an innovative technique aimed at extending the depth of focus (DOF) of optics imaging systems. In digital imaging systems, super-resolution digital reconstruction close to the diffraction limit of optical systems has always been a hot research topic. With the design of a point spread function (PSF) generated by a suitably phase mask, WFC could also be used in super-resolution image reconstruction. In this paper, we use a deep learning network combined with WFC as a general framework for images reconstruction, and verify its possibility and effectiveness. Considering the blur and additive noise simultaneously, we proposed three super-resolution image reconstruction procedures utilizing convolutional neural networks (CNN) based on mean square error (MSE) loss, conditional Generative Adversarial Networks (CGAN), and Swin Transformer Networks (SwinIR) based on mean absolute error (MAE) loss. We verified their effectiveness by simulation experiments. A comparison of experimental results shows that the SwinIR deep residual network structure based on MAE loss optimization criteria can generate more realistic super-resolution images with more details. In addition, we used a WFC camera to obtain a resolution test target and real scene images for experiments. Using the resolution test target, we demonstrated that the spatial resolution could be improved from 55.6 lp/mm to 124 lp/mm by the proposed super-resolution reconstruction procedure. The reconstruction results show that the proposed deep learning network model is superior to the traditional method in reconstructing high-frequency details and effectively suppressing noise, with the resolution approaching the diffraction limit.

Reconstructing Cancellous Bone From Down-Sampled Optical-Resolution Photoacoustic Microscopy Images With Deep Learning

ObjectiveBone diseases deteriorate the microstructure of bone tissue. Optical-resolution photoacoustic microscopy (OR-PAM) enables high spatial resolution of imaging bone tissues. However, the spatiotemporal trade-off limits the application of OR-PAM. The purpose of this study was to improve the quality of OR-PAM images without sacrificing temporal resolution. MethodsIn this study, we proposed the Photoacoustic Dense Attention U-Net (PADA U-Net) model, which was used for reconstructing full-scanning images from under-sampled images. Thereby, this approach breaks the trade-off between imaging speed and spatial resolution. ResultsThe proposed method was validated on resolution test targets and bovine cancellous bone samples to demonstrate the capability of PADA U-Net in recovering full-scanning images from under-sampled OR-PAM images. With a down-sampling ratio of [4, 1], compared to bilinear interpolation, the Peak Signal-to-Noise Ratio and Structural Similarity Index Measure values (averaged over the test set of bovine cancellous bone) of the PADA U-Net were improved by 2.325 dB and 0.117, respectively. ConclusionThe results demonstrate that the PADA U-Net model reconstructed the OR-PAM images well with different levels of sparsity. Our proposed method can further facilitate early diagnosis and treatment of bone diseases using OR-PAM.

Autofocusing in digital holography based on an adaptive genetic algorithm

In digital holography (DH), determining the reconstruction distance is critical to the quality of the reconstructed image. However, traditional focal plane detection methods require considerable time investment to reconstruct and evaluate holograms at multiple distances. To address this inefficiency, this paper proposes a fast and accurate autofocusing method based on an adaptive genetic algorithm. This method only needs to find several reconstruction distances in the search area as an initial population, and then adaptively optimize the reconstruction distance through iteration to determine the optimal focal plane in the search area. In addition, an off-axis digital holographic optical system was used to capture the holograms of the USAF resolution test target and the coin. The simulation and experimental results indicated that, compared with the traditional autofocusing, the proposed method can reduce the computation time by about 70% and improve the focal plane accuracy by up to 0.5 mm.

Direct Numerical Modeling as a Tool for Optical Coherence Tomography Development: SNR (Sensitivity) and Lateral Resolution Test Target Interpretation

Optical Coherence Tomography (OCT) is a growing family of biophotonic imaging techniques, but in the literature there is a lack of easy-to-use tools to universally directly evaluate a device’s theoretical performance for a given metric. Modern computing tools mean that direct numerical modeling can, from first principles, simulate the performance metrics of a specific device directly without relying on analytical approximations and/or complexities. Here, we present two different direct numerical models, along with the example MATLAB code for the reader to adapt to their own systems. The first model is of photo-electron shot noise at the detector, the primary noise source for OCT. We use this firstly to evaluate the amount of additional noise present (1.5 dB) for an experimental setup. Secondly, we demonstrate how to use it to precisely quantify the expected shot noise SNR limit difference between time-domain and Fourier-domain OCT systems in a given hypothetical experiment. The second model is used to demonstrate how USAF 1951 test chart images should be interpreted for a given lateral PSF shape. Direct numerical modeling is an easy and powerful basic tool for researchers and developers, the wider use of which may improve the rigor of the OCT literature.

Live Cell Imaging by Single-Shot Common-Path Wide Field-of-View Reflective Digital Holographic Microscope.

Quantitative phase imaging by digital holographic microscopy (DHM) is a nondestructive and label-free technique that has been playing an indispensable role in the fields of science, technology, and biomedical imaging. The technique is competent in imaging and analyzing label-free living cells and investigating reflective surfaces. Herein, we introduce a new configuration of a wide field-of-view single-shot common-path off-axis reflective DHM for the quantitative phase imaging of biological cells that leverages several advantages, including being less-vibration sensitive to external perturbations due to its common-path configuration, also being compact in size, simple in optical design, highly stable, and cost-effective. A detailed description of the proposed DHM system, including its optical design, working principle, and capability for phase imaging, is presented. The applications of the proposed system are demonstrated through quantitative phase imaging results obtained from the reflective surface (USAF resolution test target) as well as transparent samples (living plant cells). The proposed system could find its applications in the investigation of several biological specimens and the optical metrology of micro-surfaces.

Cepstrum-based interferometric microscopy (CIM) for quantitative phase imaging

A universal methodology for coding-decoding the complex amplitude field of an imaged sample in coherent microscopy is presented, where no restrictions on any of the two interferometric beams are required. Thus, the imaging beam can be overlapped with, in general, any other complex amplitude distribution and, in particular, with a coherent and shifted version of itself considering two orthogonal directions. The complex field values are retrieved by a novel Cepstrum-based algorithm, named as Spatial-Shifting Cepstrum (SSC), based on a weighted subtraction of the Cepstrum transform in the cross-correlation term of the object field spectrum in addition with the generation of a complex pupil from the combination of the information retrieved from different holographic recordings (one in horizontal and one in vertical direction) where one of the interferometric beams is shifted 1 pixel. As a result, the field of view is tripled since the complex amplitudes of the three interferometric fields involved in the process are retrieved. Proof-of-concept validation of this methodology, named as Cepstrum-based Interferometric Microscopy (CIM), is provided considering an off-axis holographic configuration for retrieving the cross-correlation of the two interferometric complex amplitude fields in a compact quasi-common path Michelson interferometric configuration. Experimental results for different types of phase samples (resolution test targets for step-by-step calibration and demonstration as well as fixed biosamples) are included.

Wafer‐Scale Manufacturing of Near‐Infrared Metalenses

AbstractMetalenses have revolutionized optical technologies with their superior ability to manipulate light and the potential to replace conventional, bulky optical components. However, their commercialization is hindered by limitations in conventional manufacturing techniques, such as small patterning areas, low throughput, and high cost. In this study, two methods are introduced for scalable and wafer‐scale manufacturing of metalenses operating in the near‐infrared region, aimed at overcoming the abovementioned challenges. The first type of metalens is polarization‐independent and constructed using hydrogenated amorphous silicon cylindrical structures fabricated through direct photolithography. This metalens has a diameter of 1 cm and numerical aperture (NA) of 0.53. The focusing efficiency is confirmed at a 940 nm wavelength, and the focal spot profile approaches the diffraction limit. The second metalens is polarization‐dependent and fabricated using silicon nanoparticle‐embedded‐resin rectangular structures through a cost‐effective nanoimprinting method. This process can produce metalenses with a diameter of 5 mm and an NA of 0.53. Both types of metalenses demonstrate high‐resolution capabilities when imaging a 1951 USAF resolution test target and bioimaging. This research offers innovative pathways for the mass production and large‐scale fabrication of metalenses. It is believed that the work will accelerate the industrialization of metalenses, fostering further advances in optical technology.

Multi-planar low-coherence diffraction imaging

Coherent diffraction imaging (CDI) is a popular methodology for quantitative phase restoration and wavefront measurements from several desired intensity measurements that have wide applications ranging from wavefront sensing in adaptive optics to optical imaging. To overcome the problem of image quality degradation caused by the high coherence of the light source, a multi-planar low-coherent diffraction imaging (LCDI) method under low-coherence light sources was proposed and verified by measuring an U.S. Air Force (USAF) resolution test target and a nine-letter element. Negative lenses with different focal lengths were designed as filters in the image-relay system to capture multi-planar diffraction patterns at a single recording plane, and a multi-planar LCDI algorithm was used to reconstruct the test sample using a set of intensity images. Its feasibility was demonstrated experimentally using a super-radiation light-emitting diode. Considering its compact setup, easy realization, high speed, and easy operation in phase recovery, this method is effective for rapidly detecting phase and wavefront measurements with high accuracy and resolution.

Image restoration algorithm for terahertz FMCW radar imaging.

The terahertz frequency modulation continuous-wave (THz FMCW) imaging technology has been widely used in non-destructive testing applications. However, THz FMCW real-aperture radar usually has a small depth of field and poor lateral resolution, thus restricting the high-precision imaging application. This paper proposes a 150-220GHz FMCW Bessel beam imaging system, effectively doubling the depth of field and unifying the lateral resolution compared to the Gaussian beam quasi-optical system. Moreover, a THz image restoration algorithm based on local gradients and convolution kernel priors is proposed to eliminate further the convolution effect introduced by the Bessel beam, thereby enhancing the lateral resolution to 2mm. It effectively improves the image under-restoration or over-restoration caused by the mismatch between the ideal and actual point spread function. The imaging results of the resolution test target and semiconductor device verify the advantages of the proposed system and algorithm.

A fiber-coupled laser diode design for reflection mode optical resolution photoacoustic microscopy

Pulsed Laser Diodes (PLD) are compact and high pulse repetition rate laser sources that show a great potential for low-cost Optical Resolution Photoacoustic Microscopes (OR-PAM). Nevertheless, their non-uniform multimode laser beams are of low quality so that high lateral resolutions with tightly focused beams are difficult to realize at long focusing distances, as required for reflection mode OR-PAM devices of clinical application. A new strategy based on homogenizing and shaping the laser diode beam with a square-core multimode optical fiber allowed to attain competitive lateral resolutions while keeping one centimeter working distance. The theoretical expressions for the laser spot size, determining optical lateral resolution, and the depth of focus are also written for general multimode beams. An OR-PAM system was built in confocal reflection mode with a linear phased-array as the ultrasound receiver in order to test its performance, first, on a resolution test target and, afterwards, on ex vivo rabbit ears to show the system potential for subcutaneous imaging of blood vessels and hair follicles.

Diffraction-limited hyperspectral mid-infrared micro-ellipsometry.

The recent introduction of quantum cascade lasers (QCL) in infrared spectroscopic ellipsometry led to decisive improvements in measurement times and signal-to-noise ratios of this powerful analytical method. In this contribution, we present another significant enhancement leading to the first, to the best of our knowledge, diffraction-limited micro-ellisometry setup in the mid-infrared spectral range with a spatial resolution better than 13.3 µm. The fast spectral tunability of the QCL combined with phase-modulated polarization enabled simultaneous acquisition of broadband (900 cm-1-1204 cm-1) high-resolution (1 cm-1) hyperspectral Ψ, Δ-cubes in a scanning approach in reasonable time scales. The spatial resolution of the QCL micro-ellipsometer was experimentally characterized by the knife-edge method and measurements of a resolution test target. Furthermore, the hyperspectral ellipsometric investigation of a polymer multilayer cross section and the portrait window of a 200-euro bank note demonstrate the capabilities of diffraction-limited QCL micro-ellipsometry.

Single-shot wavelength-multiplexed phase microscopy under Gabor regime in a regular microscope embodiment

Phase imaging microscopy under Gabor regime has been recently reported as an extremely simple, low cost and compact way to update a standard bright-field microscope with coherent sensing capabilities. By inserting coherent illumination in the microscope embodiment and producing a small defocus distance of the sample at the input plane, the digital sensor records an in-line Gabor hologram of the target sample, which is then numerically post-processed to finally achieve the sample’s quantitative phase information. However, the retrieved phase distribution is affected by the two well-known drawbacks when dealing with Gabor’s regime, that is, coherent noise and twin image disturbances. Here, we present a single-shot technique based on wavelength multiplexing for mitigating these two effects. A multi-illumination laser source (including 3 diode lasers) illuminates the sample and a color digital sensor (conventional RGB color camera) is used to record the wavelength-multiplexed Gabor hologram in a single exposure. The technique is completed by presenting a novel algorithm based on a modified Gerchberg–Saxton kernel to finally retrieve an enhanced quantitative phase image of the sample, enhanced in terms of coherent noise removal and twin image minimization. Experimental validations are performed in a regular Olympus BX-60 upright microscope using a 20X 0.46NA objective lens and considering static (resolution test targets) and dynamic (living spermatozoa) phase samples.

Fluholoscopy—Compact and Simple Platform Combining Fluorescence and Holographic Microscopy

The combination of different imaging modalities into single imaging platforms has a strong potential in biomedical sciences as it permits the analysis of complementary properties of the target sample. Here, we report on an extremely simple, cost-effective, and compact microscope platform for achieving simultaneous fluorescence and quantitative phase imaging modes with the capability of working in a single snapshot. It is based on the use of a single illumination wavelength to both excite the sample's fluorescence and provide coherent illumination for phase imaging. After passing the microscope layout, the two imaging paths are separated using a bandpass filter, and the two imaging modes are simultaneously obtained using two digital cameras. We first present calibration and analysis of both fluorescence and phase imaging modalities working independently and, later on, experimental validation for the proposed common-path dual-mode imaging platform considering static (resolution test targets, fluorescent micro-beads, and water-suspended lab-made cultures) as well as dynamic (flowing fluorescent beads, human sperm cells, and live specimens from lab-made cultures) samples.

High-resolution polarization-sensitive Fourier ptychography microscopy using a high numerical aperture dome illuminator.

Polarization-sensitive Fourier-ptychography microscopy (pFPM) allows for high resolution imaging while maintaining a large field of view, and without mechanical movements of optical-setup components. In contrast to ordinary light microscopes, pFPM provides quantitative absorption and phase information, for complex and birefringent specimens, with high resolution across a wide field of view. Using a semi-spherical home-built LED illumination array, a single polarizer, and a 10x /0.28NA objective, we experimentally demonstrate high performance pFPM with a synthesized NA of 1.1. Applying the standard quantitative method, a measured half-pitch resolution of 244 nm is achieved for the 1951 USAF resolution test target. As application examples, the polarimetric properties of a herbaceous flowering plant and the metastatic carcinoma of human liver cells are analyzed and quantitatively imaged.

Low-intensity illumination for lensless digital holographic microscopy with minimized sample interaction.

Exposure to laser light alters cell culture examination via optical microscopic imaging techniques based on label-free coherent digital holography. To mitigate this detrimental feature, researchers tend to use a broader spectrum and lower intensity of illumination, which can decrease the quality of holographic imaging due to lower resolution and higher noise. We study the lensless digital holographic microscopy (LDHM) ability to operate in the low photon budget (LPB) regime to enable imaging of unimpaired live cells with minimized sample interaction. Low-cost off-the-shelf components are used, promoting the usability of such a straightforward approach. We show that recording data in the LPB regime (down to 7 µW of illumination power) does not limit the contrast or resolution of the hologram phase and amplitude reconstruction compared to regular illumination. The LPB generates hardware camera shot noise, however, to be effectively minimized via numerical denoising. The ability to obtain high-quality, high-resolution optical complex field reconstruction was confirmed using the USAF 1951 amplitude sample, phase resolution test target, and finally, live glial restricted progenitor cells (as a challenging strongly absorbing and scattering biomedical sample). The proposed approach based on severely limiting the photon budget in lensless holographic microscopy method can open new avenues in high-throughout (optimal resolution, large field-of-view, and high signal-to-noise-ratio single-hologram reconstruction) cell culture imaging with minimized sample interaction.

Programmable Aperture Using a Digital Micromirror Device for In-Line Holographic Microscopy

We present the novel use of a digital micromirror device (DMD) as a digitally adjustable and programmable aperture for lens-free digital in-line holographic microscopy (LFDIHMY). The proposed micro-electro-mechanical system (MEMS) based DMD aperture can be used as a compact and inexpensive replacement for existing mechanical based aperture control methods required for many resolution enhancement methods. Simulation of the DMD aperture in an in-line holography setup is presented using a general DMD model from literature. A hardware prototype was successfully implemented and demonstrated. The resolution of both simulated and experimental holograms recorded with a conventional pinhole aperture and the proposed DMD aperture are compared using a 1951 USAF resolution test target. We show that a resolution of 2.46 <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\mathrm { \mu {\textbf {m}}}$ </tex-math></inline-formula> , close to the sensor pixel limit, can be obtained with each aperture. This demonstrates that a DMD can be used as a programmable replacement for a pinhole aperture.

Single-shot off-axis digital holographic system with extended field-of-view by using multiplexing method

We propose a new configuration of single-shot off-axis digital holographic system to realize double the camera field-of-view (FOV) of the existing off-axis Mech-Zehnder type holographic setup. The double FOV is obtained by double spatial frequency multiplexing of two different areas of an object beam by inserting a Fresnel bi-prism in it, which divides the object beam into two, both carrying different object information. The image sensor is placed at the plane where these two different FOVs overlap so as to record simultaneously two parts of the wavefront of the object in a single-shot. The multiplexed hologram is carrying two interferometric images corresponding to two different FOVs of the object which are modulated with two different spatial carrier frequencies. The feasibility of the proposed digital holographic system is experimentally demonstrated by imaging two different areas of a resolution test target. The limitation of the proposed system and a method to overcome it, are also discussed. The proposed system is useful in a wide range of applications including microscopy and optical metrology.

Fully Integrated Ultra-thin Intraoperative Micro-imager for Cancer Detection Using Upconverting Nanoparticles

PurposeIntraoperative detection and removal of microscopic residual disease (MRD) remain critical to the outcome of cancer surgeries. Today’s minimally invasive surgical procedures require miniaturization and surgical integration of highly sensitive imagers to seamlessly integrate into the modern clinical workflow. However, current intraoperative imagers remain cumbersome and still heavily dependent on large lenses and rigid filters, precluding further miniaturization and integration into surgical tools.ProceduresWe have successfully engineered a chip-scale intraoperative micro-imager array—without optical filters or lenses—integrated with lanthanide-based alloyed upconverting nanoparticles (aUCNPs) to achieve tissue imaging using a single micro-chip. This imaging platform is able to leverage the unique optical properties of aUCNPs (long luminescent lifetime, high-efficiency upconversion, no photobleaching) by utilizing a time-resolved imaging method to acquire images using a 36-by-80-pixel, 2.3 mm times 4.8 mm silicon-based electronic imager micro-chip, that is, less than 100-µm thin. Each pixel incorporates a novel architecture enabling automated background measurement and cancellation. We have validated the performance, spatial resolution, and the background cancellation scheme of the imaging platform, using resolution test targets and mouse prostate tumor sample intratumorally injected with aUCNPs. To demonstrate the ability to image MRD, or tumor margins, we evaluated the imaging platform in visualizing a single-cell thin section of the injected prostate tumor sample.ResultsTested on USAF resolution targets, the imager is able to achieve a resolution of 71 µm. We have also demonstrated successful background cancellation, achieving a signal-to-background ratio of 8 when performing ex vivo imaging on aUCNP-injected prostate tumor sample, improved from originally 0.4. The performance of the imaging platform on single-cell layer sections was also evaluated and the sensor achieved a signal-to-background ratio of 4.3 in resolving cell clusters with sizes as low as 200 cells.ConclusionThe imaging system proposed here is a scalable chip-scale ultra-thin alternative for bulky conventional intraoperative imagers. Its novel pixel architecture and background correction scheme enable visualization of microscopic-scale residual disease while remaining completely free of lenses and filters, achieving an ultra-miniaturized form factor—critical for intraoperative settings.

Tunable optofluidic microbubble lens.

Optofluidic microlenses are one of the crucial components in many miniature lab-on-chip systems. However, many optofluidic microlenses are fabricated through complex micromachining and tuned by high-precision actuators. We propose a kind of tunable optofluidic microbubble lens that is made by the fuse-and-blow method with a fiber fusion splicer. The optical focusing properties of the microlens can be tuned by changing the refractive index of the liquid inside. The focal spot size is 2.8 µm and the focal length is 13.7 µm, which are better than those of other tunable optofluidic microlenses. The imaging capability of the optofluidic microbubble lens is demonstrated under a resolution test target and the imaging resolution can reach 1 µm. The results indicate that the optofluidic microbubble lens possesses good focusing properties and imaging capability for many applications, such as cell counting, optical trapping, spatial light coupling, beam shaping and imaging.