Absorption Contrast Research Articles

Optical single-pixel volumetric imaging by three-dimensional light-field illumination

Three-dimensional single-pixel imaging (3D SPI) has become an attractive imaging modality for both biomedical research and optical sensing. 3D-SPI techniques generally depend on time-of-flight or stereovision principle to extract depth information from backscattered light. However, existing implementations for these two optical schemes are limited to surface mapping of 3D objects at depth resolutions, at best, at the millimeter level. Here, we report 3D light-field illumination single-pixel microscopy (3D-LFI-SPM) that enables volumetric imaging of microscopic objects with a near-diffraction-limit 3D optical resolution. Aimed at 3D space reconstruction, 3D-LFI-SPM optically samples the 3D Fourier spectrum by combining 3D structured light-field illumination with single-element intensity detection. We build a 3D-LFI-SPM prototype that provides an imaging volume of ∼390 × 390 × 3,800 μm3 and achieves 2.7-μm lateral resolution and better than 37-μm axial resolution. Its capability of 3D visualization of label-free optical absorption contrast is demonstrated by imaging single algal cells in vivo. Our approach opens broad perspectives for 3D SPI with potential applications in various fields, such as biomedical functional imaging.

Deformation of two-phase aggregates with in situ X-ray tomography in rotating Paris-Edinburgh cell at GPa pressures and high temperature.

High-pressure (>1 GPa) torsion apparatus can be coupled with in situ X-ray tomography (XRT) to study microstructures in materials associated with large shear strains. Here, deformation experiments were carried out on multi-phase aggregates at ∼3-5 GPa and ∼300-500°C, using a rotational tomography Paris-Edinburgh press (RoToPEc) with in situ absorption contrast XRT on the PSICHE beamline at Synchrotron SOLEIL. The actual shear strain reached in the samples was quantified with respect to the anvil twisting angles, which is γ ≤ 1 at 90° anvil twist and reaches γ ≃ 5 at 225° anvil twist. 2D and 3D quantifications based on XRT that can be used to study in situ the deformation microfabrics of two-phase aggregates at high shear strain are explored. The current limitations for investigation in real time of deformation microstructures using coupled synchrotron XRT with the RoToPEc are outlined.

Design of an agile optical switcher based on a 1D silver plasmonic nanograting filled with a nonlinear Kerr material

In this paper, we investigate the nonlinear optical response characteristics of a metallic nanograting with nonlinear Kerr media within its slits using the finite element method. The proposed nanograting system is illuminated by a nanosecond Gaussian pulse laser under normal incidence and the electric filed pointing across the slits. The results show a perfect linear absorption at resonance wavelength thanks to the coupling of the surface plasmon resonance (SPR) mode and photon cavity mode. We simulate the transient nonlinear absorption variation of the system when the pulse laser is set up at either resonance or off-resonance wavelengths. The results indicate that the unit linear absorption drastically decreases by increasing the laser fluence around the center of the pulse. Interestingly, one can also enhance the weak linear off-resonance absorption to the value of unit by increasing the pulse laser fluence. The higher the laser fluence, the higher the maximum absorption contrast between linear and nonlinear regimes occurs owing to the nonlinear Kerr effect. Indeed, when the laser fluence reaches a critical value, it can excite the Kerr nonlinearity, which changes the coupling strength of SPR mode and the photon cavity mode leading to the absorption adjustment in the nanograting. These properties indicate the possibility of utilizing the proposed nanograting in dual functional absorber and nonabsorber systems, which make it an appropriate candidate for agile optical switching devices.

Optimization of phase contrast imaging with a nano-focus x-ray tube.

Propagation-based phase contrast imaging with a laboratory x-ray source is a valuable tool for studying samples that show only low absorption contrast, either because of low density, elemental composition, or small feature size. If a propagation distance between sample and detector is introduced and the illumination is sufficiently coherent, the phase shift in the sample will cause additional contrast around interfaces, known as edge enhancement fringes. The strength of this effect depends not only on sample parameters and energy but also on the experimental geometry, which can be optimized accordingly. Recently, x-ray lab sources using transmission targets have become available, which provide very small source sizes in the few hundred nanometer range. This allows the use of a high-magnification geometry with a very short source-sample distance, while still achieving sufficient spatial coherence at the sample position. Moreover, the high geometrical magnification makes it possible to use detectors with a larger pixel size without reducing the image resolution. Here, we explore the influence of magnification on the edge enhancement fringes in such a geometry. We find experimentally and theoretically that the fringes become maximal at a magnification that is independent of the total source-detector distance. This optimal magnification only depends on the source size, the steepness of the sample feature, and the detector resolution. A stronger influence of the sample feature on the optimal magnification compared to low-magnification geometries is observed.

A deep learning reconstruction framework for low dose phase contrast computed tomography via inter-contrast enhancement

Phase contrast computed tomography (PCCT) offers excellent imaging contrast on soft tissue while it generate absorption, phase and dark-field contrast tomographic images. It has shown a great potential in clinical diagnosis. However, existing PCCT methods require high radiation doses. Reducing tube current is a universal low dose approach while it will introduce quantum noise in projections. In this paper, we report a deep learning (DL) framework for low dose PCCT based on inter-contrast enhancement. It utilizes the multi-contrast feature of PCCT and the varying effects of noise on each contrast. The missing structure in the contrasts that are more affected by noise can be recovered by those that are less affected. Considering the grating-based PCCT as example, the proposed framework is validated with experiments and a dramatic quality improvement of multi-contrast tomographic images has been obtained. This study shows potential of DL techniques in the field of low dose PCCT.

Spectroscopic Microtomography in the Short-Wave Infrared Wavelength Range.

Spectroscopic microtomography provides the ability to perform 4D (3D structural and 1D chemical) imaging of a thick microscopic specimen. Here, we demonstrate spectroscopic microtomography in the short-wave infrared (SWIR) wavelength using digital holographic tomography, which captures both the absorption coefficient and refractive index. A broadband laser in tandem with a tunable optical filter allows us to scan the wavelength from 1100 to 1650 nm. Using the developed system, we measure human hair and sea urchin embryo samples. The resolution estimated with gold nanoparticles is 1.51 μm (transverse) and 1.57 μm (axial) for the field of view of 307 × 246 μm2. The developed technique will enable accurate and efficient analyses of microscopic specimens that have a distinctive absorption or refractive index contrast in the SWIR range.

Azimuth mapping of fibrous tissue in linear dichroism-sensitive photoacoustic microscopy

Photoacoustic imaging (PAI) has emerged as a molecular-selective imaging technology based on optical absorption contrast. Dichroism-sensitive photoacoustic (DS-PA) imaging has been reported, where the absorption coefficient has a vector characteristic, featuring dimensions of contrast in polarization and wavelength. Herein, we present a DS-PA microscopy (DS-PAM) system that implements optical anisotropy contrast and molecular selectivity. Moreover, we propose mathematical solutions to fully derive dichroic properties. A wavelength for the PAI of collagenous tissue was used, and the proposed algorithms were validated using linear dichroic materials. We successfully mapped dichroic information in fibrous tissue imaging based on the degree of anisotropy and axis orientation, and also deduced mechanical assessment from the tissue arrangement. The proposed DS-PAM system and algorithms have great potential in various diagnostic fields using polarimetry, such as musculoskeletal and cardiovascular systems.

Polychromatic neutron phase-contrast imaging of weakly absorbing samples enabled by phase retrieval.

The use of a phase-retrieval technique for propagation-based phase-contrast neutron imaging with a polychromatic beam is demonstrated. This enables imaging of samples with low absorption contrast and/or improving the signal-to-noise ratio to facilitate e.g. time-resolved measurements. A metal sample, designed to be close to a phase pure object, and a bone sample with canals partially filled with D2O were used for demonstrating the technique. These samples were imaged with a polychromatic neutron beam followed by phase retrieval. For both samples the signal-to-noise ratios were significantly improved and, in the case of the bone sample, the phase retrieval allowed for separation of bone and D2O, which is important for example for in situ flow experiments. The use of deuteration contrast avoids the use of chemical contrast enhancement and makes neutron imaging an interesting complementary method to X-ray imaging of bone.

Fabrication and characterization of iodine photonic microcells for sub-Doppler spectroscopy applications.

We report on the development of all-fiber stand-alone iodine-filled photonic microcells demonstrating record absorption contrast at room temperature. The microcell's fiber is made of inhibited coupling guiding hollow-core photonic crystal fibers. The fiber-core loading with iodine was undertaken at 10-1-10-2mbar vapor pressure using what, to the best of our knowledge, is a novel gas-manifold based on metallic vacuum parts with ceramic coated inner surfaces for corrosion resistance. The fiber is then sealed on the tips and mounted on FC/APC connectors for better integration with standard fiber components. The stand-alone microcells display Doppler lines with contrasts up to 73% in the 633 nm wavelength range, and an off-resonance insertion loss between 3 to 4 dB. Sub-Doppler spectroscopy based on saturable absorption has been carried out to resolve the hyperfine structure of the P(33)6-3 lines at room temperature with a full-width at half maximum of 24 MHz on the b4 component with the help of lock-in amplification. Also, we demonstrate distinguishable hyperfine components on the R(39)6-3 line at room temperature without any recourse to signal-to-noise ratio amplification techniques.

Location-Dependent Spatiotemporal Antialiasing in Photoacoustic Computed Tomography.

Photoacoustic computed tomography (PACT) images optical absorption contrast by detecting ultrasonic waves induced by optical energy deposition in materials such as biological tissues. An ultrasonic transducer array or its scanning equivalent is used to detect ultrasonic waves. The spatial distribution of the transducer elements must satisfy the spatial Nyquist criterion; otherwise, spatial aliasing occurs and causes artifacts in reconstructed images. The spatial Nyquist criterion poses different requirements on the transducer elements' distributions for different locations in the image domain, which has not been studied previously. In this research, we elaborate on the location dependency through spatiotemporal analysis and propose a location-dependent spatiotemporal antialiasing method. By applying this method to PACT in full-ring array geometry, we effectively mitigate aliasing artifacts with minimal effects on image resolution in both numerical simulations and in vivo experiments.

Photoacoustic Imaging and Characterization of Bone in Medicine: Overview, Applications, and Outlook.

Photoacoustic techniques have shown promise in identifying molecular changes in bone tissue and visualizing tissue microstructure. This capability represents significant advantages over gold standards (i.e., dual-energy X-ray absorptiometry) for bone evaluation without requiring ionizing radiation. Instead, photoacoustic imaging uses light to penetrate through bone, followed by acoustic pressure generation, resulting in highly sensitive optical absorption contrast in deep biological tissues. This review covers multiple bone-related photoacoustic imaging contributions to clinical applications, spanning bone cancer, joint pathologies, spinal disorders, osteoporosis, bone-related surgical guidance, consolidation monitoring, and transsphenoidal and transcranial imaging. We also present a summary of photoacoustic-based techniques for characterizing biomechanical properties of bone, including temperature, guided waves, spectral parameters, and spectroscopy. We conclude with a future outlook based on the current state of technological developments, recent achievements, and possible new directions.

Photoacoustic imaging with limited sampling: a review of machine learning approaches.

Photoacoustic imaging combines high optical absorption contrast and deep acoustic penetration, and can reveal structural, molecular, and functional information about biological tissue non-invasively. Due to practical restrictions, photoacoustic imaging systems often face various challenges, such as complex system configuration, long imaging time, and/or less-than-ideal image quality, which collectively hinder their clinical application. Machine learning has been applied to improve photoacoustic imaging and mitigate the otherwise strict requirements in system setup and data acquisition. In contrast to the previous reviews of learned methods in photoacoustic computed tomography (PACT), this review focuses on the application of machine learning approaches to address the limited spatial sampling problems in photoacoustic imaging, specifically the limited view and undersampling issues. We summarize the relevant PACT works based on their training data, workflow, and model architecture. Notably, we also introduce the recent limited sampling works on the other major implementation of photoacoustic imaging, i.e., photoacoustic microscopy (PAM). With machine learning-based processing, photoacoustic imaging can achieve improved image quality with modest spatial sampling, presenting great potential for low-cost and user-friendly clinical applications.

Full-field quantitative X-ray phase nanotomography via space-domain Kramers–Kronig relations

Given the low absorption contrast of X-rays, phase shift has been playing an important role as an alternative source of contrast in X-ray nanoimaging. Numerous phase-measuring techniques have been proposed, most of which, however, are based on significant assumptions or sample translations. In this study, we propose the application of Kramers–Kronig (KK) relations in the spatial domain as a solution to allow the X-ray quantitative phase image to be directly calculated from the measured intensity image without any additional requirements. Based on this straightforward principle, we have presented KK nanotomography by introducing a spatial-frequency cutoff filter into a conventional tomographic setup. The robustness and versatility of the proposed method were experimentally verified based on various sample tomograms. We expect KK nanotomography to be widely adopted as a powerful and easy-to-adapt phase quantification solution for X-ray microscopes.

Photoacoustic Imaging of COVID-19 Vaccine Site Inflammation of Autoimmune Disease Patients.

Based on the observations made in rheumatology clinics, autoimmune disease (AD) patients on immunosuppressive (IS) medications have variable vaccine site inflammation responses, whose study may help predict the long-term efficacy of the vaccine in this at-risk population. However, the quantitative assessment of the inflammation of the vaccine site is technically challenging. In this study analyzing AD patients on IS medications and normal control subjects, we imaged the inflammation of the vaccine site 24 h after mRNA COVID-19 vaccinations were administered using both the emerging photoacoustic imaging (PAI) method and the established Doppler ultrasound (US) method. A total of 15 subjects were involved, including 6 AD patients on IS and 9 normal control subjects, and the results from the two groups were compared. Compared to the results obtained from the control subjects, the AD patients on IS medications showed statistically significant reductions in vaccine site inflammation, indicating that immunosuppressed AD patients also experience local inflammation after mRNA vaccination but not in as clinically apparent of a manner when compared to non-immunosuppressed non-AD individuals. Both PAI and Doppler US were able to detect mRNA COVID-19 vaccine-induced local inflammation. PAI, based on the optical absorption contrast, shows better sensitivity in assessing and quantifying the spatially distributed inflammation in soft tissues at the vaccine site.

A versatile laboratory setup for high resolution X-ray phase contrast tomography and scintillator characterization.

X-ray micro-tomography (μCT) is a powerful non-destructive 3D imaging method applied in many scientific fields. In combination with propagation-based phase-contrast, the method is suitable for samples with low absorption contrast. Phase contrast tomography has become available in the lab with the ongoing development of micro-focused tube sources, but it requires sensitive and high-resolution X-ray detectors. The development of novel scintillation detectors, particularly for microscopy, requires more flexibility than available in commercial tomography systems. We aim to develop a compact, flexible, and versatile μCT laboratory setup that combines absorption and phase contrast imaging as well as the option to use it for scintillator characterization. Here, we present details on the design and implementation of the setup. We used the setup for μCT in absorption and propagation-based phase-contrast mode, as well as to study a perovskite scintillator. We show the 2D and 3D performance in absorption and phase contrast mode, as well as how the setup can be used for testing new scintillator materials in a realistic imaging environment. A spatial resolution of around 1.3μm is measured in 2D and 3D. The setup meets the needs for common absorption μCT applications and offers increased contrast in phase contrast mode. The availability of a versatile laboratory μCT setup allows not only for easy access to tomographic measurements, but also enables a prompt monitoring and feedback beneficial for advances in scintillator fabrication.

Virtual differential phase-contrast and dark-field imaging of x-ray absorption images via deep learning.

Weak absorption contrast in biological tissues has hindered x-ray computed tomography from accessing biological structures. Recently, grating-based imaging has emerged as a promising solution to biological low-contrast imaging, providing complementary and previously unavailable structural information of the specimen. Although it has been successfully applied to work with conventional x-ray sources, grating-based imaging is time-consuming and requires a sophisticated experimental setup. In this work, we demonstrate that a deep convolutional neural network trained with a generative adversarial network can directly convert x-ray absorption images into differential phase-contrast and dark-field images that are comparable to those obtained at both a synchrotron beamline and a laboratory facility. By smearing back all of the virtual projections, high-quality tomographic images of biological test specimens deliver the differential phase-contrast- and dark-field-like contrast and quantitative information, broadening the horizon of x-ray image contrast generation.

Micro-Computed Tomography Beamline of the Australian Synchrotron: Micron-Size Spatial Resolution X-ray Imaging

The first new beamline of the BRIGHT project—involving the construction of eight new beamlines at the Australian Synchrotron—is the Micro-Computed Tomography (MCT) beamline. MCT will extend the facility’s capability for higher spatial resolution X-ray-computed tomographic imaging allowing for commensurately smaller samples in comparison with the existing Imaging and Medical Beamline (IMBL). The source is a bending-magnet and it is operating in the X-ray energy range from 8 to 40 keV. The beamline provides important new capability for a range of biological and material-science applications. Several imaging modes will be offered such as various X-ray phase-contrast modalities (propagation-based, grating-based, and speckle-based), in addition to conventional absorption contrast. The unique properties of synchrotron radiation sources (high coherence, energy tunability, and high brightness) are predominantly well-suited for producing phase contrast data. An update on the progress of the MCT project in delivering high-spatial-resolution imaging (in the order of micron size) of mm-scale objects will be presented in detail with some imaging results from the hot-commissioning stage.

High-fidelity deep functional photoacoustic tomography enhanced by virtual point sources

Photoacoustic tomography (PAT), a hybrid imaging modality that acoustically detects the optical absorption contrast, is a promising technology for imaging hemodynamic functions in deep tissues far beyond the traditional optical microscopy. However, the most clinically compatible PAT often suffers from the poor image fidelity, mostly due to the limited detection view of the linear ultrasound transducer array. PAT can be improved by employing highly-absorbing contrast agents such as droplets and nanoparticles, which, however, have low clinical translation potential due to safety concerns and regulatory hurdles imposed by these agents. In this work, we have developed a new methodology that can fundamentally improve PAT’s image fidelity without hampering any of its functional capability or clinical translation potential. By using clinically-approved microbubbles as virtual point sources that strongly and isotropically scatter the local pressure waves generated by surrounding hemoglobin, we can overcome the limited-detection-view problem and achieve high-fidelity functional PAT in deep tissues, a technology referred to as virtual-point-source PAT (VPS-PAT). We have thoroughly investigated the working principle of VPS-PAT by numerical simulations and in vitro phantom experiments, clearly showing the signal origin of VPSs and the resultant superior image fidelity over traditional PAT. We have also demonstrated in vivo applications of VPT-PAT for functional small-animal studies with physiological challenges. We expect that VPS-PAT can find broad applications in biomedical research and accelerated translation to clinical impact.

Refractive index profiling of multimode specialty optical fibers by absorption contrast X-ray computed microtomography

We report on successful refractive index profiling of commercially available step-index and in-house made graded-index multimode specialty optical fibers by means of X-ray computed microtomography. Our results demonstrate that the latter is an advantageous technique for characterizing large core optical fibers, which allows for retrieving information about the refractive index at optical frequencies by exploiting the absorption coefficient of X-rays.

Simulation Study of Sub-Period Super-Resolution X-ray Phase Imaging with Triangular Phase Grating

X-ray phase contrast image has higher sensitivity to materials with light elements than the conventional absorption contrast. Phase image measurement technique using diffraction gratings can be used in the laboratory. Furthermore, X-ray phase microscope using the diffraction gratings has been also developed. However, phase measurement with diffraction gratings has a drawback that the spatial resolution is limited to the grating period. To overcome this problem, a scanning system using a triangular phase grating has recently been proposed, which is expected to archive a large field of view and a high spatial resolution. This study aims to investigate the feasibility of the system and a deep understanding of the optical system by simulation.