Differential phase contrast for quantitative imaging and spectro-microscopy at a nanoprobe beamline

Microsphere-assisted imaging has been demonstrated to improve imaging resolution and features a simple operation, low cost, and compatibility with multiple microscopic imaging techniques. In this Letter, we present an imaging method that integrates microsphere-assisted imaging with the differential phase contrast (DPC) technique. The DPC images correlated with phase gradients were generated by capturing the imaging results of microsphere-assisted imaging under asymmetric illumination. Imaging experiments using Blu-ray disc stripes demonstrated that the DPC technique can enhance the contrast and optimize the imaging quality of microsphere-assisted nano-imaging.

In this work we present the results of differential phase contrast (DPC) [1,2] measurements of micro magnetic field distributions in annealed cobalt thin films. The polycrystalline specimens with a thickness of about 50 nm were produced by thermal boat evaporation under high vacuum conditions. After annealing the average size of individual cobalt crystals is increased and their magnetic properties are changed as shown by G. Herzer in [3] and [4]. Our goal was to determine, if DPC is a suitable technique to investigate how these changes in the crystallographic structure effect the magnetic properties of our specimen. Therefore we performed single DPC measurements as well as DPC tilting series to investigate if and how the magnetic behaviour of the annealed cobalt films is changed. We investigated 5 different samples. One being the untempered thin film and the others were annealed over one hour with increasing temperatures ranging from 520 K to 820 K. To investigate the magnetic properties we performed a DPC tilt series for each sample. By tilting the specimen relative to the magnetic field of the objective lens we change the effective external in plane field on our samples. This leads to a change of the in plane induction in our cobalt thin films. We started each tilting series at large angles of α=20°, to be sure that all magnetic moments in our specimen are aligned in one direction. Figure 1 shows a DPC measurement of such a saturated state. The homogeneous green colour of the measurement confirms that the magnetic induction is saturated along the direction indicated by the colourwheel. Figures 2‐4 show an excerpt of DPC measurements performed during the tilt series of the specimen annealed at 850 K. It can be seen, that the magnetic structure changes with decreasing α and it seems, that one of the magnetic ripples is pinned to the highlighted crystallites in the center of figures 2 and 3. By calculating the average direction and strength of the beam deflection for each individual DPC measurement we get information about the dependency between the magnetic induction and the tilting angle respectively the external magnetic field applied. We will present the results of our measurements on the different annealed specimen showing hysteretic behaviour of the magnetic induction and discuss the differences between them. During our experiments especially the tilt series, which take about 4 hours to obtain, we encountered some difficulties with the DPC technique. In this work we will present these shortcomings and show solutions to some of the problems occured.

Quantitative differential phase contrast (DPC) microscopy has been developed to retrieval the phase of object leveraged from weak object assumption. In perspective of transfer function of DPC, deviating from that with coherent illumination, the DPC reveals non-isotropic transfer function under partial coherent illumination. Therefore, multi-axis measurements would be necessary to formulate the isotropic response for quantitative DPC phase imaging. In this paper, the gradient amplitude modulation is used to generate the isotropic DPC transfer function with only two-axis measurements. We show our modeling and simulation results.

Differential Phase Contrast (DPC) microscopy is a practical method for recovering quantitative phase from intensity images captured with different source patterns in an LED array microscope. Being a partially coherent imaging method, DPC does not suffer from speckle artifacts and achieves 2× better resolution than coherent methods. Like all imaging systems, however, DPC is susceptible to aberrations. Here, we propose a method of algorithmic self-calibration for DPC where we simultaneously recover the complex-field of the sample and the spatially-variant aberrations of the system, using 4 images with different illumination source patterns. The resulting phase reconstructions are digitally aberration-corrected.

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.

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 300 μm. 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).

Purpose: The anatomical noise power spectra (NPS) for differential phase contrast (DPC) and dark field (DF) imaging have recently been characterized using a power-law model with two parameters, alpha and beta, an innovative extension to the methodology used in x-ray attenuation based breast imaging such as mammography, DBT, or cone-beam CT. Beta values of 3.6, 2.6, and 1.3 have been measured for absorption, DPC, and DF respectively for cadaver breasts imaged in the coronal plane; these dramatic differences should be reflected in their detection performance. The purpose of this study was to determine the impact of anatomical noise on breast calcification detection and compare the detection performance of the three contrast mechanisms of a multi-contrast x-ray imaging system. Methods: In our studies, a calcification image object was segmented out of the multi-contrast images of a cadaver breast specimen. 50 measured total NPS were measured from breast cadavers directly. The ideal model observer detectability was calculated for a range of doses (5–100%) and a range of calcification sizes (diameter = 0.25–2.5 mm). Results: Overall we found the highest average detectability corresponded to DPC imaging (7.4 for 1 mm calc.), with DF the next highest (3.8 for 1 mm calc.), and absorption the lowest (3.2 for 1 mm calc.). However, absorption imaging also showed the slowest dependence on dose of the three modalities due to the significant anatomical noise. DPC showed a peak detectability for calcifications ∼1.25 mm in diameter, DF showed a peak for calcifications around 0.75 mm in diameter, and absorption imaging had no such peak in the range explored. Conclusion: Understanding imaging performance for DPC and DF is critical to transition these modalities to the clinic. The results presented here offer new insight into how these modalities complement absorption imaging to maximize the likelihood of detecting early breast cancers. J. Garrett, Y. Ge, K. Li: Nothing to disclose. G.-H. Chen: Research funded, GE Healthcare; Research funded, Siemens AX.

We describe a simple and fast technique to perform ultrasound differential phase contrast (DPC) imaging in arbitrarily thick scattering media. Although configured in a reflection geometry, DPC is based on transmission imaging and is a direct analog of optical differential interference contrast. DPC exploits the memory effect and works in combination with standard pulse-echo imaging, with no additional hardware or data requirements, enabling complementary phase contrast (in the transverse direction) without any need for intensive numerical computation. We experimentally demonstrate the principle of DPC using tissue phantoms with calibrated speed-of-sound inclusions.

Introducing a non-pixelated and fast centre of mass detector for differential phase contrast microscopy

Differential phase contrast (DPC) is a non-interferometric quantitative phase imaging method achieved by using an asymmetric imaging procedure. We report a pupil modulation differential phase contrast (PMDPC) imaging method by filtering a sample's Fourier domain with half-circle pupils. A phase gradient image is captured with each half-circle pupil, and a quantitative high resolution phase image is obtained after a deconvolution process with a minimum of two phase gradient images. Here, we introduce PMDPC quantitative phase image reconstruction algorithm and realize it experimentally in a 4f system with an SLM placed at the pupil plane. In our current experimental setup with the numerical aperture of 0.36, we obtain a quantitative phase image with a resolution of 1.73μm after computationally removing system aberrations and refocusing. We also extend the depth of field digitally by 20 times to ±50μm with a resolution of 1.76μm.

We introduce a lock-in method to increase the phase contrast in incoherent Differential Phase Contrast (DPC) imaging. The use of a smart pixel detector with in-pixel signal demodulation, paired with synchronized illumination, provides the basis of a bit-efficient approach to DPC. The experiments show an increased sensitivity by a factor of 8, for equivalent standard DPC measurements; single-shot sensitivity of 0.7 mrad at a frame rate of 1400 fps is demonstrated. This new approach may open the way for the use of incoherent phase microscopy in biological applications where extreme phase sensitivity and millisecond response time is required.

Quantitative differential phase contrast (qDPC) microscopy enables high‐resolution, label‐free imaging of weakly absorbing samples by combining asymmetrical illumination with phase transfer function (PTF) deconvolution. However, conventional methods are limited by the ill‐posed nature of deconvolution and the band‐limited characteristics of the PTF, leading to poor robustness against noise and background fluctuations, particularly in thick or complex samples. To overcome these challenges, we propose a pupil‐driven differential phase contrast (PD‐DPC) framework that integrates system PTFs into both the data fidelity and regularization terms of the reconstruction model. The proposed model incorporates an edge‐sparsity‐promoting regularization to preserve structural detail and suppress noise, along with a Retinex‐inspired fidelity formulation to mitigate background fluctuations. The resulting non‐convex optimization problem is solved via an efficient Split Bregman algorithm with iterative reweighted soft‐thresholding. Simulations and experiments demonstrate that PD‐DPC outperforms L2‐DPC, Iso‐DPC, TV‐DPC, and Retinex‐DPC in terms of background suppression, phase fidelity, and edge preservation. The framework is compatible with diverse DPC modalities and enables automatic cell contour segmentation as well as high‐resolution imaging of absorbing tissues beyond the weak‐object approximation. By combining physics‐informed priors with a data‐adaptive reconstruction strategy, PD‐DPC offers a robust, broadly applicable solution that substantially enhances the accuracy and applicability of qDPC for biomedical imaging. The MATLAB code is available on GitHub .

In this work, non-Lorentz, Lorentz transmission electron microscopy (LTEM) and differential phase contrast (DPC) methods were used to observe the phase and magnetic structure of the alnico magnet. All observations were performed on alnico-5 in transverse orientation. The results showed the magnetic contrast of the domain walls in the conventional TEM images despite the effects of the lens magnetic field, which was verified and explained using DPC. Experimental methodology and sample results are summarized with advice for further research.

Quantitative differential phase-contrast (DPC) microscopy produces phase images of transparent objects based on a number of intensity images. To reconstruct the phase, in DPC microscopy, a linearized model for weakly scattering objects is considered; this limits the range of objects to be imaged, and requires additional measurements and complicated algorithms to correct for system aberrations. Here, we present a self-calibrated DPC microscope using an untrained neural network (UNN), which incorporates the nonlinear image formation model. Our method alleviates the restrictions on the object to be imaged and simultaneously reconstructs the complex object information and aberrations, without any training dataset. We demonstrate the viability of UNN-DPC microscopy through both numerical simulations and LED microscope-based experiments.

Cellphones equipped with high-quality cameras and powerful CPUs as well as GPUs are widespread. This opens new prospects to use such existing computational and imaging resources to perform medical diagnosis in developing countries at a very low cost. Many relevant samples, like biological cells or waterborn parasites, are almost fully transparent. As they do not exhibit absorption, but alter the light’s phase only, they are almost invisible in brightfield microscopy. Expensive equipment and procedures for microscopic contrasting or sample staining often are not available. Dedicated illumination approaches, tailored to the sample under investigation help to boost the contrast. This is achieved by a programmable illumination source, which also allows to measure the phase gradient using the differential phase contrast (DPC) [1, 2] or even the quantitative phase using the derived qDPC approach [3]. By applying machine-learning techniques, such as a convolutional neural network (CNN), it is possible to learn a relationship between samples to be examined and its optimal light source shapes, in order to increase e.g. phase contrast, from a given dataset to enable real-time applications. For the experimental setup, we developed a 3D-printed smartphone microscope for less than 100 $ using off-the-shelf components only such as a low-cost video projector. The fully automated system assures true Koehler illumination with an LCD as the condenser aperture and a reversed smartphone lens as the microscope objective. We show that the effect of a varied light source shape, using the pre-trained CNN, does not only improve the phase contrast, but also the impression of an improvement in optical resolution without adding any special optics, as demonstrated by measurements.