Full-field Microscopy Research Articles

Laboratory X-ray Microscopy of 3D Nanostructures in the Hard X-ray Regime Enabled by a Combination of Multilayer X-ray Optics.

High-resolution imaging of buried metal interconnect structures in advanced microelectronic products with full-field X-ray microscopy is demonstrated in the hard X-ray regime, i.e., at photon energies > 10 keV. The combination of two multilayer optics-a side-by-side Montel (or nested Kirkpatrick-Baez) condenser optic and a high aspect-ratio multilayer Laue lens-results in an asymmetric optical path in the transmission X-ray microscope. This optics arrangement allows the imaging of 3D nanostructures in opaque objects at a photon energy of 24.2 keV (In-Kα X-ray line). Using a Siemens star test pattern with a minimal feature size of 150 nm, it was proven that features < 150 nm can be resolved. In-Kα radiation is generated from a Ga-In alloy target using a laboratory X-ray source that employs the liquid-metal-jet technology. Since the penetration depth of X-rays into the samples is significantly larger compared to 8 keV photons used in state-of-the-art laboratory X-ray microscopes (Cu-Kα radiation), 3D-nanopattered materials and structures can be imaged nondestructively in mm to cm thick samples. This means that destructive de-processing, thinning or cross-sectioning of the samples are not needed for the visualization of interconnect structures in microelectronic products manufactured using advanced packaging technologies. The application of laboratory transmission X-ray microscopy in the hard X-ray regime is demonstrated for Cu/Cu6Sn5/Cu microbump interconnects fabricated using solid-liquid interdiffusion (SLID) bonding.

Propagation-based phase-contrast imaging method for full-field X-ray microscopy using advanced Kirkpatrick-Baez mirrors.

We demonstrate a propagation-based phase-contrast imaging method for full-field X-ray microscopy based on advanced Kirkpatrick-Baez (AKB) mirrors to achieve high-contrast observations of weak phase objects and correct field curvature aberrations. Through a demonstration performed at SPring-8, the phase contrast of weak phase objects such as polystyrene spheres and chemically fixed cells was successfully observed with high sensitivity (∼0.03 rad). Furthermore, the field of view of the AKB mirrors was expanded to the full area of the obtained images (25 × 30 µm) by correcting the field curvature aberration using reconstructed complex wavefields.

Live-cell analysis framework for quantitative phase imaging with slightly off-axis digital holographic microscopy

Label-free quantitative phase imaging is an essential tool for studying in vitro living cells in various research fields of life sciences. Digital holographic microscopy (DHM) is a non-destructive full-field microscopy technique that provides phase images by directly measuring the optical path differences, which facilitates cell segmentation and allows the determination of several important cellular physical features, such as dry mass. In this work, we present a systematic analysis framework for live-cell imaging and morphological characterization, terms as LAF (live-cell analysis framework). All image processing algorithms involved in this framework are implemented on the high-resolution artifact-free quantitative phase images obtained by our previously proposed slightly off-axis holographic system (FPDH) and associated reconstruction methods. A highly robust automated cell segmentation method is applied to extract the valid cellular region, followed by live-cell analysis framework algorithms to determine the physical and morphological properties, including the area, perimeter, irregularity, volume and dry mass, of each individual cell. Experiments on live HeLa cells demonstrate the validity and effectiveness of the presented framework, revealing its potential for diverse biomedical applications.

Label-free full-field Doppler phase microscopy based on optical computation.

The capability to image subtle mechanical motion at cellular and sub-cellular scales can be used to study how extracellular particles interact with cultured cells and, more generally, how cells interact with their environment. However, current technologies need to provide sufficient spatial resolution, temporal resolution, and motion sensitivity to image cellular and sub-cellular motion in the en face plane. To address this unmet need, we investigate a full-field Doppler phase microscopy (FF-DPM) technology based on an innovative optical computation strategy that enables depth-resolved imaging and phase quantification. In this study, we validated the motion tracking (displacements and velocities) capability of FF-DPM by imaging samples actuated by a piezo transducer (PZT). We demonstrated FF-DPM imaging of magnetic particles under different conditions with different motion characteristics. Our results show that free particles (suspended in a cell culture medium) had a significantly larger magnitude of motion than particles adhered to a cell. The key innovation of this study is the use of an optical computation strategy to perform depth-resolved phase quantification and Doppler measurement. The FF-DPM will have a significant impact, as it provides a unique capability to quantitatively measure subtle motion for models based on cultured cells.

The Projection and Transmission X-ray Microscopy at Taiwan Photon Source

Projection X-ray microscopy (PXM) and transmission X-ray microscopy (TXM) have been established at beamline 31 at the Taiwan Photon Source (TPS). The TPS 31A beamline is sourced by a wiggler with multiple beam modes: white beam, high-flux mono-beam, and high-resolution mono-beam modes. The PXM is based on a collimated beam, and the TXM is full-field microscopy based on a zoneplate. The PXM system and TXM share the same detector platform, which is equipped with different kinds of detector systems for PXM and TXM. The PXM was designed for high-speed 3D tomography with an automatic tray system for loading and unloading samples, including a robotic arm and pre-alignment system. Both the PXM and TXM systems were designed for use in situ, which requires longer working distance for the sample’s environmental cell.

Full-field chromatic confocal microscopy for surface profilometry with sub-micrometer accuracy

In various manufacturing processes like semiconductor fabrication, highly accurate and efficient surface profilometry on microstructures is critically demanded. This article presents a quasi-area-scan chromatic confocal microscopy for full-field surface profilometry with high precision and accuracy for in-line automated optical inspection (AOI). Unlike conventional confocal microscopy, the proposed opto-mechatronic confocal measuring system does not require mechanical scanning of the test object. Moreover, it is bi-telecentric to satisfy full-field measurement without optical aberration. Optical line-scan illumination and imaging modules are integrated with an electrically driven galvanometer to perform line scanning. Most importantly, the proposed system is superior to traditional chromatic confocal scanning in that positioning uncertainties can be avoided, and high-speed detection with a large field of view and high accuracy can be achieved. Pre-calibrated step-height gauges were experimentally measured to verify the precision and accuracy of the developed probe. Results showed measurement bias kept below 0.239 µm with a standard deviation of 0.126 µm. When the Galvanometer reaches its design specifications, the measuring speed of the system can exceed 8000 lines/s.

X-ray directional dark-field imaging using Unified Modulated Pattern Analysis.

X-ray directional dark-field imaging is a recent technique that can reveal a sample’s small-scale structural properties which are otherwise invisible in a conventional imaging system. In particular, directional dark-field can detect and quantify the orientation of anisotropic structures. Here, we present an algorithm that allows for the extraction of a directional dark-field signal from X-ray speckle-based imaging data. The experimental setup is simple, as it requires only the addition of a diffuser to a full-field microscope setup. Sandpaper is an appropriate diffuser material in the hard x-ray regime. We propose an approach to extract the mean scattering width, directionality, and orientation from the recorded speckle images acquired with the technique. We demonstrate that our method can detect and quantify the orientation of fibres inside a carbon fibre reinforced polymer (CFRP) sample within one degree of accuracy and show how the accuracy depends on the number of included measurements. We show that the reconstruction parameters can be tuned to increase or decrease accuracy at the expense of spatial resolution.

Nanoscale Antiferromagnetic Domain Imaging using Full-Field Resonant X-ray Magnetic Diffraction Microscopy.

The physical properties of magnetic materials frequently depend not only on the microscopic spin and electronic structures, but also on the structures of mesoscopic length scales that emerge, for instance, from domain formations, or chemical and/or electronic phase separations. However, experimental access to such mesoscopic structures is currently limited, especially for antiferromagnets with net zero magnetization. Here, full-field microscopy and resonant magnetic X-ray diffraction are combined to visualize antiferromagnetic (AF) domains of the spin-orbit Mott insulator Sr2 IrO4 with area over ≈0.1 mm2 and with spatial resolution as high as ≈150nm. With the unprecedented wide field of views and high spatial resolution, an intertwining of two AF domains on a length comparable to the measured average AF domain wall width of 545nm is revealed. This mesoscopic structure comprises a substantial portion of the sample surface, and thus can result in a macroscopic response unexpected from its microscopic magnetic structure. In particular, the symmetry analysis presented in this work shows that the inversion symmetry, which is preserved by the microscopic AF order, becomes ill-defined at the mesoscopic length scale. This result underscores the importance of this novel technique for a thorough understanding of the physical properties of antiferromagnets.

In situ imaging of temperature-dependent fast and reversible nanoscale domain switching in a single-crystal perovskite

Metal halide perovskites exhibit a rich crystal structure, with multiple phases as well as ferroelastic domains, which is crucial for the optical and electrical properties. The average crystal phase-transition temperatures can be shifted by size, strain, or defects, but it is not clear whether such differences can also appear locally within a single crystal. The experimental study of domain dynamics within nanocrystals is challenging and requires a method capable of probing crystal lattice variations with both high spatial and temporal resolution. Here, we show that in situ full-field diffraction x-ray microscopy can be used to image domains in a single crystal ${\mathrm{CsPbBr}}_{3}$ nanoplatelet as the temperature traverses the orthorhombic to tetragonal phase transition, at 150 nm spatial resolution and 6 s time resolution. The images reveal sudden domain pattern changes faster than the temporal resolution. Surprisingly, we observe substantial local variations during heating, with domain changes occurring at different temperatures within the single crystal. The nanoplatelet exhibits a high-temperature domain pattern completely different from the low-temperature one, but both patterns are reproducible, and we reversibly switch between them in multiple cycles. These results demonstrate that single ${\mathrm{CsPbBr}}_{3}$ crystals can exhibit substantial local variation of their basic crystal properties.

Optimizing the energy bandwidth for transmission full-field X-ray microscopy experiments.

Full-field transmission X-ray microscopy (TXM) is a very potent high-resolution X-ray imaging technique. However, it is challenging to achieve fast acquisitions because of the limited efficiency of the optics. Using a broader energy bandwidth, for example using a multilayer monochromator, directly increases the flux in the experiment. The advantage of more counts needs to be weighed against a deterioration in achievable resolution because focusing optics show chromatic aberrations. This study presents theoretical considerations of how much the resolution is affected by an increase in bandwidth as well as measurements at different energy bandwidths (ΔE/E = 0.013%, 0.27%, 0.63%) and the impact on achievable resolution. It is shown that using a multilayer monochromator instead of a classical silicon double-crystal monochromator can increase the flux by an order of magnitude with only a limited effect on theresolution.

Partial-field illumination ophthalmoscope: improving the contrast of a camera-based retinal imager.

Effective and accurate in vivo diagnosis of retinal pathologies requires high performance imaging devices, combining a large field of view and the ability to discriminate the ballistic signal from the diffuse background in order to provide a highly contrasted image of the retinal structures. Here, we have implemented the partial-field illumination ophthalmoscope, a patterned illumination modality, integrated to a high pixel rate adaptive optics full-field microscope. This non-invasive technique enables us to mitigate the low signal-to-noise ratio, intrinsic of full-field ophthalmoscopes, by partially illuminating the retina with complementary patterns to reconstruct a wide-field image. This new, to the best of our knowledge, modality provides an image contrast spanning from the full-field to the confocal contrast, depending on the pattern size. As a result, it offers various trade-offs in terms of contrast and acquisition speed, guiding the users towards the most efficient system for a particular clinical application.

Stochasticity in the Switching of Nanodisks for Probabilistic Computing

Author(s): Han, HS; Lee, S; Je, SG; Kang, M; Ok, HJ; Kim, N; Chao, W; Im, MY; Lee, KS | Abstract: Stochasticity in magnetic nanodevices is an essential characteristic for harnessing these devices to computing based on population coding or the building blocks of probabilistic computing, p-bits. A magnetic tunneling junction (MTJ) consisting of a patterned magnetic element is considered a promising computing unit in the concept of artificial neurons and p-bits. A comprehensive understanding of the stochasticity in the switching of patterned magnetic elements is crucial for realizing MTJ-based probabilistic computing technology. In the present work, the stochastic behavior in the switching process of a perpendicularly magnetized Co/Pt disk within an array was directly observed utilizing full-field soft X-ray microscopy. Within 50 repeated hysteretic cycles, the stochastic magnetization switching of individual Co/Pt disks within disk arrays is identified. We found that the stochasticity in the magnetization switching of disks considerably depends on the disk size. The stochasticity initially decreases as the disk radius gets bigger from 125 to 375 nm (region I), then increases with further enlarging the disk size to 625 nm (region II). The variance of thermal fluctuation relevant to the disk size and the multilevel switching within a disk are severely involved in the observed size-dependent stochasticity. This work provides the way for controlling the stochasticity in the switching of nanopatterned elements, which is a key aspect of MTJ-based probabilistic computing.

Multi-Lens Array Full-Field X-ray Microscopy

X-ray full-field microscopy at laboratory sources for photon energies above 10 keV suffers from either long exposure times or low resolution. The photon flux is mainly limited by the objectives used, having a limited numerical aperture NA. We show that this can be overcome by making use of the cone-beam illumination of laboratory sources by imaging the same field of view (FoV) several times under slightly different angles using an array of X-ray lenses. Using this technique, the exposure time can be reduced drastically without any loss in terms of resolution. A proof-of-principle is given using an existing laboratory metal-jet source at the 9.25 keV Ga Kα-line and compared to a ray-tracing simulation of the setup.

De-scattering with Excitation Patterning enables rapid wide-field imaging through scattering media.

Nonlinear optical microscopy has enabled in vivo deep tissue imaging on the millimeter scale. A key unmet challenge is its limited throughput especially compared to rapid wide-field modalities that are used ubiquitously in thin specimens. Wide-field imaging methods in tissue specimens have found successes in optically cleared tissues and at shallower depths, but the scattering of emission photons in thick turbid samples severely degrades image quality at the camera. To address this challenge, we introduce a novel technique called De-scattering with Excitation Patterning or "DEEP," which uses patterned nonlinear excitation followed by computational imaging-assisted wide-field detection. Multiphoton temporal focusing allows high-resolution excitation patterns to be projected deep inside specimen at multiple scattering lengths due to the use of long wavelength light. Computational reconstruction allows high-resolution structural features to be reconstructed from tens to hundreds of DEEP images instead of millions of point-scanning measurements.

High-energy x-ray nanotomography introducing an apodization Fresnel zone plate objective lens.

In this study, high-energy x-ray nanotomography (nano-computed tomography, nano-CT) based on full-field x-ray microscopy was developed. Fine two-dimensional and three-dimensional (3D) structures with linewidths of 75 nm-100nm were successfully resolved in the x-ray energy range of 15 keV-37.7 keV. The effective field of view was ∼60 µm, and the typical measurement time for one tomographic scan was 30 min-60min. The optical system was established at the 250-m-long beamline 20XU of SPring-8 to realize greater than 100× magnification images. An apodization Fresnel zone plate (A-FZP), specifically developed for high-energy x-ray imaging, was used as the objective lens. The design of the A-FZP for high-energy imaging is discussed, and its diffraction efficiency distribution is evaluated. The spatial resolutions of this system at energies of 15 keV, 20 keV, 30 keV, and 37.7 keV were examined using a test object, and the measured values are shown to be in good agreement with theoretical values. High-energy x-ray nano-CT in combination with x-ray micro-CT is applied for 3D multiscale imaging. The entire bodies of bulky samples, ∼1mm in diameter, were measured with the micro-CT, and the nano-CT was used for nondestructive observation of regions of interest. Examples of multiscale CT measurements involving carbon steel, mouse bones, and a meteorite are discussed.

Analysis of the impact of optical aberrations in en-face full-field OCT microscopy.

Optical coherence tomography (OCT) is a powerful technique for cross-sectioning imaging. However, the lateral resolution may be degraded by optical aberrations originating from the sample or the setup. We present an extensive quantitative study of the impact of aberrations in time-domain en-face full-field OCT (FFOCT). Using an adaptive optics loop integrated in an FFOCT setup, a deformable mirror is used to introduce low-order calibrated aberrations. The experimental analysis of both the line spread functions (SF) and the complex object images has allowed us to measure the loss in contrast and the impact on lateral spatial resolution. We demonstrate that the frequency content of FFOCT image spectra in terms of signal-to-noise ratio and cutoff frequency is degraded by aberrations but remains much higher than in conventional incoherent images. Line SF profiles in conventional imaging display widening, whereas in FFOCT they display oscillations, leading to the possible perception of preserved resolution. Nevertheless, for complex objects, the aberration image blurring is strong due to the convolution process by the point SF, resulting in a significant filtering of the image spatial spectrum.

Biological Applications of Short Wavelength Microscopy Based on Compact, Laser-Produced Gas-Puff Plasma Source

Over the last decades, remarkable efforts have been made to improve the resolution in photon-based microscopes. The employment of compact sources based on table-top laser-produced soft X-ray (SXR) in the “water window” spectral range (λ = 2.3–4.4 nm) and extreme ultraviolet (EUV) plasma allowed to overcome the limitations imposed by large facilities, such as synchrotrons and X-ray free electron lasers (XFEL), because of their high complexity, costs, and limited user access. A laser-plasma double stream gas-puff target source represents a powerful tool for microscopy operating in transmission mode, significantly improving the spatial resolution into the nanometric scale, comparing to the traditional visible light (optical) microscopes. Such an approach allows generating the plasma efficiently, without debris, providing a high flux of EUV and SXR photons. In this review, we present the development and optimization of desktop imaging systems: a EUV and an SXR full field microscope, allowing to achieve a sub-50 nm spatial resolution with short exposure time and an SXR contact microscope, capable to resolve internal structures in a thin layer of sensitive photoresist. Details about the source, as well as imaging results for biological applications, will be presented and discussed.

Thermal generation, manipulation and thermoelectric detection of skyrmions

Recent years have witnessed significant progresses in realizing skyrmions in chiral magnets1-4 and asymmetric magnetic multilayers5-13, as well as their electrical manipulation2,7,8,10. Equally important, thermal generation, manipulation and detection of skyrmions can be exploited for prototypical new architecture with integrated computation14 and energy harvesting15. It has yet to verify if skyrmions can be purely generated by heating16,17, and if their resultant direction of motion driven by temperature gradients follows the diffusion or, oppositely, the magnonic spin torque17-21. Here, we address these important issues in microstructured devices made of multilayers: (Ta_CoFeB_MgO)15, (Pt_CoFeB_MgO_Ta)15 and (Pt_Co_Ta)15 integrated with on-chip heaters, by using a full-field soft X-ray microscopy. The thermal generation of densely packed skyrmions is attributed to the low energy barrier at the device edge, together with the thermally induced morphological transition from stripe domains to skyrmions. The unidirectional diffusion of skyrmions from the hot region towards the cold region is experimentally observed. It can be theoretically explained by the combined contribution from repulsive forces between skyrmions, and thermal spin-orbit torques in competing with magnonic spin torques17,18,20,21 and entropic forces22. These thermally generated skyrmions can be further electrically detected by measuring the accompanied anomalous Nernst voltages23. The on-chip thermoelectric generation, manipulation and detection of skyrmions could open another exciting avenue for enabling skyrmionics, and promote interdisciplinary studies among spin caloritronics15, magnonics24 and skyrmionics3,4,12.

Recent progress in synchrotron radiation 3D-4D nano-imaging based on X-ray full-field microscopy.

The advent of high-flux, high-brilliance synchrotron radiation (SR) has prompted the development of high-resolution X-ray imaging techniques such as full-field microscopy, holography, coherent diffraction imaging and ptychography. These techniques have strong potential to establish non-destructive three- and four-dimensional nano-imaging when combined with computed tomography (CT), called nano-tomography (nano-CT). X-ray nano-CTs based on full-field microscopy are now routinely available and widely used. Here we discuss the current status and some applications of nano-CT using a Fresnel zone plate as an objective. Optical properties of full-field microscopy, such as spatial resolution and off-axis aberration, which determine the effective field of view, are also discussed, especially in relation to 3D tomographic imaging.

Compact full-field hard x-ray microscope based on advanced Kirkpatrick–Baez mirrors

X-ray full-field microscopy is a promising method for nondestructive observation of opaque materials because it can attain a high resolution and wide field of view without sample scanning. We recently developed hard x-ray objective optics, which are key devices for full-field microscopy, based on total-reflection mirrors with high throughput and achromatic properties. The objective optics consist of two types of advanced Kirkpatrick–Baez mirrors configured as crossed one-dimensional Wolter type I and type III optics. The designed optics possessed magnification factors of 42–45 with a compact camera length of approximately 2 m. The hard x-ray full-field microscope based on this system was tested at the BL29XU beamline at SPring-8. We were able to resolve 100-nm periods (50-nm line widths) of a resolution test chart at a photon energy of 15 keV over 30 h, which demonstrated the remarkable stability of this system. The image quality was preserved over a wide photon energy range from 9 to 15 keV. A periodic dot pattern with dot diameters of 300 nm, formed on a 775-µm-thick Si substrate, was three-dimensionally visualized by computed tomography.