Differential Phase Contrast Imaging Research Articles

DFT-Assisted Investigation of the Electric Field and Charge Density Distribution of Pristine and Defective 2D WSe2 by Differential Phase Contrast Imaging.

Most properties of solid materials are defined by their internal electric field and charge density distributions which so far are difficult to measure with high spatial resolution. Especially for 2D materials, the atomic electric fields influence the optoelectronic properties. In this study, the atomic-scale electric field and charge density distribution of WSe2 bi- and trilayers are revealed using an emerging microscopy technique, differential phase contrast (DPC) imaging in scanning transmission electron microscopy (STEM). For pristine material, a higher positive charge density located at the selenium atomic columns compared to the tungsten atomic columns is obtained and tentatively explained by a coherent scattering effect. Furthermore, the change in the electric field distribution induced by a missing selenium atomic column is investigated. A characteristic electric field distribution in the vicinity of the defect with locally reduced magnitudes compared to the pristine lattice is observed. This effect is accompanied by a considerable inward relaxation of the surrounding lattice, which according to first principles DFT calculation is fully compatible with a missing column of Se atoms. This shows that DPC imaging, as an electric field sensitive technique, provides additional and remarkable information to the otherwise only structural analysis obtained with conventional STEM imaging.

Noise correction in differential phase contrast for improving phase sensitivity

Differential phase contrast (DPC) imaging relies on computational analysis to extract quantitative phase information from phase gradient images. However, even modest noise level can introduce errors that propagate through the computational process, degrading the quality of the final phase result and further reducing phase sensitivity. Here, we introduce the noise-corrected DPC (ncDPC) to enhance phase sensitivity. This approach is based on a theoretical DPC model that effectively considers most relevant noise sources in the camera and non-uniform illumination in DPC. In particular, the dominating shot noise and readout noise variance can be jointly estimated using frequency analysis and further corrected by block-matching 3D (BM3D) method. Finally, the denoised images are used for phase retrieval based on the common Tikhonov inversion. Our results, based on both simulated and experimental data, demonstrate that ncDPC outperforms the traditional DPC (tDPC), enabling significant improvements in both phase reconstruction quality and phase sensitivity. Besides, we have demonstrated the broad applicability of ncDPC by showing its performance in various experimental datasets.

Poling induced changes in polarization domain structure of polymer ferroelectrics for application in organic thin film transistors

While electrical poling of organic ferroelectrics has been shown to improve device properties, there are challenges in visualizing accompanying structural changes. We observe poling induced changes in ferroelectric domains by applying differential phase contrast (DPC) imaging in the scanning transmission electron microscope, a method that has been used to observe spatial distributions of electromagnetic fields at the atomic scale. In this work, we obtain DPC images from unpoled and electrically poled polyvinylidene fluoride trifluorethylene films and compare their performance in polymer thin film transistors. The vertically poled films show uniform domains throughout the bulk compared to the unpoled film with a significantly higher magnitude of the overall polarization. Thin film transistors comprising a donor–acceptor copolymer as the active semiconductor layer show improved performance with the vertically poled ferroelectric dielectric film compared with the unpoled ferroelectric dielectric film. A poling field of 80–100 MV/m for the dielectric layer yields the best performing transistors; higher than 100 MV/m is seen to degrade the transistor performance. The results are consistent with a reduction in deleterious charge carrier scattering from ferroelectric domain boundaries or interfacial dipoles arising from electrical poling.

Transport-of-intensity model for single-mask x-ray differential phase contrast imaging

X-ray phase contrast imaging holds great promise for improving the visibility of light-element materials such as soft tissues and tumors. The single-mask differential phase contrast imaging method stands out as a simple and effective approach to yield differential phase contrast. In this work, we introduce a model for a single-mask phase imaging system based on the transport-of-intensity equation. Our model provides an accessible understanding of signal and contrast formation in single-mask x-ray phase imaging, offering a clear perspective on the image formation process, for example, the origin of alternate bright and dark fringes in phase contrast intensity images. Aided by our model, we present an efficient retrieval method that yields differential phase contrast imagery in a single acquisition step. Our model gives insight into the contrast generation and its dependence on the system geometry and imaging parameters in both the initial intensity image as well as retrieved images. The model validity as well as the proposed retrieval method are demonstrated via both experimental results on a system developed in house as well as Monte Carlo simulations. In conclusion, our work not only provides a model for an intuitive visualization of image formation but also offers a method to optimize differential phase imaging setups, holding tremendous promise for advancing medical diagnostics and other applications.

A four-grating interferometer for x-ray multi-contrast imaging.

X-ray multi-contrast imaging with gratings provides a practical method to detect differential phase and dark-field contrast images in addition to the x-ray absorption image traditionally obtained in laboratory or hospital environments. Systems have been developed for preclinical applications in areas including breast imaging, lung imaging, rheumatoid arthritis hand imaging and kidney stone imaging. Prevailing x-ray interferometers for multi-contrast imaging include Talbot-Lau interferometers and universal moiré effect-based phase-grating interferometers. Talbot-Lau interferometers suffer from conflict between high interferometer sensitivity and large field of view (FOV) of the object being imaged. A small period analyzer grating is necessary to simultaneously achieve high sensitivity and large FOV within a compact imaging system but is technically challenging to produce for high x-ray energies. Phase-grating interferometers suffer from an intrinsic fringe period ranging from a few micrometers to several hundred micrometers that can hardly be resolved by large area flat panel x-ray detectors. The purpose of this work is to introduce a four-grating x-ray interferometer that simultaneously allows high sensitivity and large FOV, without the need for a small period analyzer grating. The four-grating interferometer consists of a source grating placed downstream of and close to the x-ray source, a pair of phase gratings separated by a fixed distance placed downstream of the source grating, and an analyzer grating placed upstream of and close to the x-ray detector. The object to be imaged is placed upstream of and close to the phase-grating pair. The distance between the source grating and the phase-grating pair is designed to be far larger than that between the phase-grating pair and the analyzer grating to promote simultaneously high sensitivity and large FOV. The method was evaluated by constructing a four-grating interferometer with an 8µm period source grating, a pair of phase gratings of 2.4µm period, and an 8µm period analyzer grating. The fringe visibility of the four-grating interferometer was measured to be ≈24% at 40kV and ≈18% at 50kV x-ray tube operating voltage. A quartz bead of 6mm diameter was imaged to compare the theoretical and experimental phase contrast signal with good agreement. Kidney stone specimens were imaged to demonstrate the potential of such a system for classification of kidney stones. The proposed four-grating interferometer geometry enables a compact x-ray multi-contrast imaging system with simultaneously high sensitivity and large FOV. Relaxation of the requirement for a small period analyzer grating makes it particularly suitable for high x-ray energy applications such as abdomen and chest imaging.

Disulfiram Inhibits Opsonin-Independent Phagocytosis and Migration of Human Long-Lived In Vitro Cultured Phagocytes from Multiple Inflammatory Diseases.

Disulfiram (DSF), an anti-alcoholism medicine, exerts treatment effects in patients suffering from persistent Borreliosis and also exhibits anti-cancer effects through its copper chelating derivatives and induction of oxidative stress in mitochondria. Since chronic/persistent borreliosis is characterized by increased amounts of pro-inflammatory macrophages, this study investigated opsonin-independent phagocytosis, migration, and surface marker expression of in vivo activated and in vitro cultured human monocyte-derived phagocytes (macrophages and dendritic cells) with and without DSF treatment. Phagocytosis of non-opsonized Dynabeads® M-450 and migration of macrophages and dendritic cells were monitored using live cell analyzer Juli™ Br for 24 h, imaging every 3.5 min. To simultaneously monitor phagocyte function, results were analyzed by a newly developed software based on the differential phase contrast images of cells before and after ingestion of Dynabeads. DSF decreased the phagocytic capacities exhibited by in vitro enriched and long-lived phagocytes. Although no chemotactic gradient was applied to the test system, vigorous spontaneous migration was observed. We therefore set up an algorithm to monitor and quantify both phagocytosis and migration simultaneously. DSF not only reduced phagocytosis in a majority of these long-lived phagocytes but also impaired their migration. Despite these selective effects by DSF, we found that DSF reduced the expression densities of surface antigens CD45 and CD14 in all of our long-lived phagocytes. In cells with a high metabolic activity and high mitochondrial contents, DSF led to cell death corresponding to mitochondrial oxidative stress, whereas metabolically inactive phagocytes survived our DSF treatment protocol. In conclusion, DSF affects the viability of metabolically active phagocytes by inducing mitochondrial stress and secondly attenuates phagocytosis and migration in some long-lived phagocytes.

Theory and verification of moiré fringes for x-ray three-phase grating interferometer

Dual-phase and three-phase grating x-ray interference is a promising new technique for grating-based x-ray differential phase contrast imaging. Dual-phase grating interferometers have been relatively completely studied and discussed. In this paper, the corresponding imaging fringe formula of the three-phase grating interferometer is provided. At the same time, the similarities and differences between the three-phase grating interferometer and the dual-phase grating interferometer are investigated and verified, and that the three-phase grating interferometer can produce large-period moiré fringes without using the analyzing grating is demonstrated experimentally. Finally, a simple method of designing three-phase grating and multi-grating imaging systems from geometric optics based on the thin-lens theory of gratings is presented. These theoretical formulas and experimental results provide optimization tools for designing three-phase grating interferometer systems.

Disproportionation chemistry in K 2 PtCl 4 visualized at atomic resolution using scanning transmission electron microscopy

The direct observation of a solid-state chemical reaction can reveal otherwise hidden mechanisms that control the reaction kinetics. However, probing the chemical bond breaking and formation at the molecular level remains challenging because of the insufficient spatial-temporal resolution and composition analysis of available characterization methods. Using atomic-resolution differential phase-contrast imaging in scanning transmission electron microscopy, we have visualized the decomposition chemistry of K 2 PtCl 4 to identify its transient intermediate phases and their interfaces that characterize the chemical reduction process. The crystalline structure of K 2 PtCl 4 is found to undergo a disproportionation reaction to form K 2 PtCl 6 , followed by gradual reduction to crystalline Pt metal and KCl. By directly imaging different Pt─Cl bond configurations and comparing them to models predicted via density functional theory calculations, a causal connection between the initial and final states of a chemical reaction is established, showcasing new opportunities to resolve reaction pathways through atomistic experimental visualization.

Imaging built-in electric fields and light matter by Fourier-precession TEM

We report the precise measurement of electric fields in nanostructures, and high-contrast imaging of soft matter at ultralow electron doses by transmission electron microscopy (TEM). In particular, a versatile method based on the theorem of reciprocity is introduced to enable differential phase contrast imaging and ptychography in conventional, plane-wave illumination TEM. This is realised by a series of TEM images acquired under different tilts, thereby introducing the sampling rate in reciprocal space as a tuneable parameter, in contrast to momentum-resolved scanning techniques. First, the electric field of a p–n junction in GaAs is imaged. Second, low-dose, in-focus ptychographic and DPC characterisation of Kagome pores in weakly scattering covalent organic frameworks is demonstrated by using a precessing electron beam in combination with a direct electron detector. The approach offers utmost flexibility to record relevant spatial frequencies selectively, while acquisition times and dose requirements are significantly reduced compared to the 4D-STEM counterpart.

Quantitative real-time phase microscopy for extended depth-of-field imaging based on the 3D single-shot differential phase contrast (ssDPC) imaging method.

Optical diffraction tomography (ODT) is a promising label-free imaging method capable of quantitatively measuring the three-dimensional (3D) refractive index distribution of transparent samples. In recent years, partially coherent ODT (PC-ODT) has attracted increasing attention due to its system simplicity and absence of laser speckle noise. Quantitative phase imaging (QPI) technologies represented by Fourier ptychographic microscopy (FPM), differential phase contrast (DPC) imaging and intensity diffraction tomography (IDT) need to collect several or hundreds of intensity images, which usually introduce motion artifacts when shooting fast-moving targets, leading to a decrease in image quality. Hence, a quantitative real-time phase microscopy (qRPM) for extended depth of field (DOF) imaging based on 3D single-shot differential phase contrast (ssDPC) imaging method is proposed in this research study. qRPM incorporates a microlens array (MLA) to simultaneously collect spatial information and angular information. In subsequent optical information processing, a deconvolution method is used to obtain intensity stacks under different illumination angles in a raw light field image. Importing the obtained intensity stack into the 3D DPC imaging model is able to finally obtain the 3D refractive index distribution. The captured four-dimensional light field information enables the reconstruction of 3D information in a single snapshot and extending the DOF of qRPM. The imaging capability of the proposed qRPM system is experimental verified on different samples, achieve single-exposure 3D label-free imaging with an extended DOF for 160 µm which is nearly 30 times higher than the traditional microscope system.

Simple implementation of aperture modulation quantitative differential phase contrast imaging

As one of the quantitative phase imaging methods, aperture modulation quantitative differential phase contrast (amqDPC) imaging has isotropic lateral resolution, being widely used in many fields such as biological cell morphology detection. However, the devices required to implement it are poorly controlled and very expensive. In addition, due to the weak phase approximation, there is a large axial phase error, and the full spectrum center pass during aperture modulation will cause distortion. Therefore, a method that enables simple and low-cost implementation of amqDPC, high axial phase accuracy, and isotopic lateral resolution is reported. Through simulations and experimental verifications, axial accuracy is greatly improved in the phase range of [0, π], and an isotropic lateral resolution of 870 nm can be achieved. In addition, experiments on transparent glass circular step sample and mouse kidney tissue slice also prove the practicability of this method. According to this study, researchers can accurately and simply measure the axial height of a slightly thick sample, which is achieved by capturing two intensity images to reconstruct a quantitative phase.

Van der Waals Epitaxy, Superlubricity, and Polarization of the 2D Ferroelectric SnS.

Tin monosulfide (SnS) is a two-dimensional layered semiconductor that exhibits in-plane ferroelectric order at very small thicknesses and is of interest in highly scaled devices. Here we report the epitaxial growth of SnS on hexagonal boron nitride (hBN) using a pulsed metal-organic chemical vapor deposition process. Lattice matching is observed between the SnS(100) and hBN{11̅0} planes, with no evidence of strain. Atomic force microscopy reveals superlubricity along the commensurate direction of the SnS/hBN interface, and first-principles calculations suggest that friction is controlled by the edges of the SnS islands, rather than interface interactions. Differential phase contrast imaging detects remnant polarization in SnS islands with domains that are not dictated by step-edges in the SnS. The growth of ferroelectric SnS on high quality hBN substrates is a promising step toward electrically switchable ferroelectric semiconducting devices.

A novel fusion method for X-ray phase contrast imaging based on fast adaptive bidimensional empirical mode decomposition.

X-ray phase contrast imaging (XPCI) can separate the attenuation, refraction, and scattering signals of the object. The application of image fusion enables the concentration of distinctive information into a single image. Some methods have been applied in XPCI field, but wavelet-based decomposition approaches often result in loss of original data. To explore the application value of a novel image fusion method for XPCI system and computed tomography (CT) system. The means of fast adaptive bidimensional empirical mode decomposition (FABEMD) is considered for image decomposition to avoid unnecessary information loss. A parameter δ is proposed to guide the fusion of bidimensional intrinsic mode functions which contain high-frequency information, using a pulse coupled neural network with morphological gradients (MGPCNN). The residual images are fused by the energy attribute fusion strategy. Image preprocessing and enhancement are performed on the result to ensure its quality. The effectiveness of other image fusion methods has been compared, such as discrete wavelet transforms and anisotropic diffusion fusion. The δ-guided FABEMD-MGPCNN method achieved either the first or second position in objective evaluation metrics with biological samples, as compared to other image fusion methods. Moreover, comparisons are made with other fusion methods used for XPCI. Finally, the proposed method applied in CT show expected results to retain the feature information. The proposed δ-guided FABEMD-MGPCNN method shows potential feasibility and superiority over traditional and recent image fusion methods for X-ray differential phase contrast imaging and computed tomography systems.

Tri-directional x-ray phase contrast multimodal imaging using one hexagonal mesh modulator.

Objective. X-ray phase contrast imaging is a promising technique for future clinical diagnostic as it can provide enhanced contrast in soft tissues compared to traditional x-ray attenuation-contrast imaging. However, the strict requirements on the x-ray coherence and the precise alignment of optical elements limit its applications towards clinical use. To solve this problem, mesh-based x-ray phase contrast imaging method with one hexagonal mesh is proposed for easy alignment and better image visualization.Approach. The mesh produces structured illuminations and the detector captures its distortions to reconstruct the absorption, differential phase contrast (DPC) and dark-field (DF) images of the sample. In this work, we fabricated a hexagonal mesh to simultaneously retrieve DPC and DF signals in three different directions with single shot. A phase retrieval algorithm to obtain artifacts-free phase from DPC images with three different directions is put forward and false color dark-field image is also reconstructed with tri-directional images. Mesh-shifting method based on this hexagonal mesh modulator is also proposed to reconstruct images with better image quality at the expense of increased dose.Main results. In numerical simulations, the proposed hexagonal mesh outperforms the traditional square mesh in image evaluation metrics performance and false color visualization with the same radiation dose. The experimental results demonstrate its feasiblity in real imaging systems and its advantages in quantitive imaging and better visualization. The proposed hexagonal mesh is easy to fabricate and can be successfully applied to x-ray source with it spot size up to 300μm.Significance. This work opens new possibilities for quantitative x-ray non-destructive imaging and may also be instructive for research fields such as x-ray structured illumination microscopy (SIM), x-ray spectral imaging and x-ray phase contrast and dark-field computed tomography (CT).

In vivo optical trapping of erythrocytes in mouse liver imaged with oblique back-illumination microscopy

Optical tweezers are ideal tools for cell manipulation in vivo due to their non-contact trapping ability. In the current studies, the optical trapping of cells can only be achieved in light-permeable organs. It is important to expand the application of optical tweezers to opaque tissues and organs, where biological activities are the primary focus of biomedical research. However, the optical manipulation of cells in these opaque organs cannot be imaged using a conventional transillumination microscope due to the opacity of these organs. Here, we use optical tweezers to trap erythrocytes and measure the cell deformability in mouse liver, which are imaged with oblique back-illumination microscopy (OBM). In the microscope system, two fibers are fixed at the same oblique angle on both sides of the microscopic objective, illuminating the trapping target asymmetrically. The dual-wavelength OBM strategy allows for obtaining a differential phase contrast image in a single-shot by color channel separation. The OBM can image the flowing and trapped erythrocytes in vivo in real-time. Furthermore, the erythrocyte deformability in vivo is evaluated with optical tweezers. The spring stiffness in vivo is 8.2 ± 2.1 μN m−1, which is obtained by stretching the trapped cells with blood drag flow. Our works make it possible to utilize optical tweezers to study the live cell dynamics in opaque organs, which will expand the application of optical tweezers in the fields of biomechanical studies.

Structural Complexity and Loss of Long-range Order in θ-Al2O3 as Revealed by HAADF and Differential Phase Contrast Imaging.

Structural Complexity and Loss of Long-range Order in θ-Al2O3 as Revealed by HAADF and Differential Phase Contrast Imaging.

3D Sectioning of Rough Interfaces Using Mixed-State Multislice Ptychography, Annular Dark Field, and Integrated Differential Phase Contrast Imaging.

3D Sectioning of Rough Interfaces Using Mixed-State Multislice Ptychography, Annular Dark Field, and Integrated Differential Phase Contrast Imaging.

STEM Tomography of Biological Samples Using Integrated Differential Phase Contrast Imaging Method.

STEM Tomography of Biological Samples Using Integrated Differential Phase Contrast Imaging Method.

Single-shot quantitative differential phase contrast imaging combined with programmable polarization multiplexing illumination.

We propose a single-shot quantitative differential phase contrast method with polarization multiplexing illumination. In the illumination module of our system, a programmable LED array is divided into four quadrants and covered with polarizing films of four different polarization angles. We use a polarization camera with polarizers before the pixels in the imaging module. By matching the polarization angle between the polarizing films over the custom LED array and the polarizers in the camera, two sets of asymmetric illumination acquisition images can be calculated from a single-shot acquisition image. Combined with the phase transfer function, we can calculate the quantitative phase of the sample. We present the design, implementation, and experimental image data demonstrating the ability of our method to obtain quantitative phase images of a phase resolution target, as well as Hela cells.

Single-shot quantitative phase contrast imaging based on deep learning.

Quantitative differential phase-contrast (DPC) imaging is one of the commonly used methods for phase retrieval. However, quantitative DPC imaging requires several pairwise intensity measurements, which makes it difficult to monitor living cells in real-time. In this study, we present a single-shot quantitative DPC imaging method based on the combination of deep learning (DL) and color-encoded illumination. Our goal is to train a model that can generate an isotropic quantitative phase image (i.e., target) directly from a single-shot intensity measurement (i.e., input). The target phase image was reconstructed using a linear-gradient pupil with two-axis measurements, and the model input was the measured color intensities obtained from a radially asymmetric color-encoded illumination pattern. The DL-based model was trained, validated, and tested using thirteen different cell lines. The total number of training, validation, and testing images was 264 (10 cells), 10 (1 cell), and 40 (2 cells), respectively. Our results show that the DL-based phase images are visually similar to the ground-truth phase images and have a high structural similarity index (>0.98). Moreover, the phase difference between the ground-truth and DL-based phase images was smaller than 13%. Our study shows the feasibility of using DL to generate quantitative phase imaging from a single-shot intensity measurement.