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Annular dark-field imaging

Annular dark-field imaging, often referred to as or high-angle annular dark-field (HAADF) imaging, is a (STEM) technique that forms images by collecting incoherently scattered electrons at high scattering angles from a focused electron probe raster-scanned across a thin specimen. This method utilizes an annular detector positioned to capture diffuse events, typically with inner collection angles around 50 milliradians and outer angles up to 200 milliradians, excluding low-angle elastically scattered or unscattered electrons that would contribute to bright-field images. The resulting images exhibit Z-contrast, where intensity is approximately proportional to Z^{1.4} to Z^2 (with Z being the ), making heavier elements appear brighter and enabling straightforward atomic-number-sensitive visualization without the complications of coherent interference effects. Developed as part of advancements in instrumentation, annular dark-field imaging provides incoherent that simplifies interpretation compared to phase-contrast methods, achieving resolution down to approximately 0.05 on aberration-corrected instruments. It is particularly valuable for applications, such as mapping compositional variations in alloys, semiconductors, and nanostructures at the scale, and can be combined with techniques like electron energy-loss spectroscopy (EELS) for site-specific . Variations like low-angle annular dark-field (LAADF) extend its utility to lighter elements and defects by using lower collection angles, providing strong . Overall, this mode has become a cornerstone of high-resolution for probing structures and defects in diverse fields including , sensors, and materials.

Fundamentals

Definition and Principles

Annular dark-field (ADF) imaging is a technique employed in () that utilizes an annular detector to gather incoherently scattered electrons, enabling the formation of images based on the intensity of these scattered electrons rather than the direct beam. This approach contrasts with bright-field imaging, which captures the unscattered or low-angle transmitted beam to produce phase-contrast images sensitive to effects. In ADF imaging, the annular detector is positioned to exclude the central direct beam, focusing instead on electrons deflected by interactions with the sample, which results in dark regions for vacuum areas and brighter signals from atomic structures. The fundamental operating principle involves raster-scanning a finely focused across the specimen surface, with the probe typically achieving sub-angstrom dimensions to enable atomic-scale . As the probe illuminates each point, electrons interact with the sample atoms through processes, primarily and thermal diffuse , and those deflected to angles greater than approximately 5° are collected by the surrounding annular detector. The resulting image intensity at each scan position corresponds directly to the flux of these high-angle scattered electrons, providing a map of the sample's structural features without reliance on coherent . This incoherent imaging mode arises because the large collection angles of the annular detector average out relationships among the scattered electrons, effectively destroying transverse and simplifying image interpretation compared to coherent techniques. Consequently, the image contrast is determined solely by the localized probability, often yielding a Z-contrast effect where intensity scales with . ADF imaging requires a dedicated STEM configuration, including aberration-corrected electron optics to form the high-resolution focused probe and precise control systems for the raster scanning pattern, ensuring uniform illumination and signal collection across the field of view.

Contrast Mechanisms

In annular dark-field (ADF) imaging, the primary contrast mechanism arises from high-angle Rutherford scattering of the incident electron probe by atomic nuclei, where the differential cross-section is given by \sigma \propto \frac{Z^2}{\sin^4(\theta/2)}, with Z denoting the atomic number and \theta the scattering angle. This angular dependence favors collection of electrons scattered at large angles, resulting in image intensity that scales approximately as I \propto Z^2 \cdot f(\theta), where f(\theta) accounts for the detector's angular acceptance function. Consequently, this produces Z-contrast, in which heavier atomic columns appear brighter due to their enhanced scattering efficiency, enabling direct visualization of atomic composition without phase contrast artifacts. A significant contribution to the incoherent signal in ADF imaging comes from (TDS), where lattice vibrations displace atoms, randomizing the phase of scattered electrons and distributing diffusely at high angles. TDS dominates the collected signal, particularly at elevated specimen temperatures where populations increase, or for heavy atoms with larger cross-sections, as the TDS also follows \propto Z^2 from underlying Rutherford processes. This incoherence ensures that image contrast reflects local atomic density rather than crystalline orientation effects, though requires accounting for temperature-dependent contributions. The probe convergence semi-angle and detector collection semi-angle critically influence contrast transfer by determining the degree of scattering angle selectivity and probe-sample interaction . A larger probe convergence angle broadens the angular range of incident electrons, enhancing overlap in multiple scattering events and promoting incoherence for improved Z-contrast fidelity, while the collection semi-angle defines the inner and outer limits that filter low-angle coherent beams from high-angle incoherent ones. Optimal settings, such as collection angles exceeding 30 mrad, minimize interference and maximize sensitivity to atomic-number variations. The annular detector briefly supports this by confining detection to an annular around the optic , isolating the desired high-angle scatters.

Historical Development

Origins in STEM

Annular dark-field (ADF) imaging emerged in the 1970s as a natural extension of the scanning transmission electron microscope (), which was invented by Albert Crewe and his team at the in 1966 to achieve high-resolution imaging of individual atoms using a focused probe. Early dark-field concepts in were developed to collect scattered electrons, with initial proposals for high-angle detection appearing in 1973 by C.J. Humphreys and collaborators, who recognized the potential for enhanced contrast from electrons scattered at larger angles to distinguish atomic species. These origins built on Crewe's foundational work, shifting from conventional bright-field imaging to dark-field modes that prioritized scattered signals for improved visibility of heavy atoms in thin specimens. The first implementations of ADF imaging employed simple annular detectors positioned below the sample to capture diffusely scattered electrons, bypassing the need for objective apertures that limited conventional (TEM) dark-field techniques. In TEM dark-field, typically involves selecting specific diffracted beams with an aperture in the back focal plane, which can introduce coherent artifacts and restrict ; in contrast, STEM's annular detectors integrated over a range of scattering angles directly in the far-field, enabling incoherent collection without such constraints. This design facilitated the direct observation of single atoms on thin carbon films, demonstrating atomic without the limitations inherent to coherent TEM . Key early publications in the emphasized ADF's incoherent properties, which mitigated the contrast issues plaguing conventional TEM by treating the scattered electrons as an incoherent superposition, akin to classical optical . For instance, , , and Langmore's 1970 work detailed how high-angle in STEM produced interpretable images of isolated heavy atoms, establishing ADF as a tool for bypassing coherent effects that obscured weak signals in TEM. These studies highlighted the technique's reliance on thermal diffuse for contrast, allowing straightforward interpretation of atomic positions and compositions. Early experiments also revealed Z-contrast behavior, where image intensity scaled approximately with the square of , providing a direct visual map of material composition. The conceptual foundations of ADF imaging drew heavily from scattering theories, particularly , which describes the interaction between incident electrons and nuclei at high angles. This framework, originally developed for alpha-particle scattering in the early , was adapted in the to model electron-nucleus interactions in , predicting that high-angle electrons follow classical trajectories dominated by unscreened nuclear potentials. By applying these theories, researchers quantified the incoherent nature of ADF signals, where multiple events averaged out information, yielding robust intensity proportional to atomic density and number.

Key Advancements

In the late and early , the development of high-angle annular dark-field () in () introduced Z-contrast , which provides direct atomic-number sensitivity and incoherent for atomic-resolution visualization of crystalline materials. Pioneered by J. Pennycook and collaborators, this technique utilized annular detectors to collect high-angle scattered electrons, enabling the of atomic columns in semiconductors and interfaces without phase artifacts, achieving resolutions approaching 0.2 nm. These advancements, detailed in foundational works from 1989 to 1992, established HAADF as a cornerstone for materials characterization by suppressing effects and enhancing chemical . The 2000s marked a transformative era with the implementation of aberration correction in , dramatically reducing probe sizes to sub-angstrom levels and elevating imaging resolution. Ondrej L. Krivanek and colleagues at Nion developed sextupole-based correctors for , compensating for and enabling beam diameters below 0.5 at 200 kV, as demonstrated in 2002 experiments resolving silicon atomic spacings at 0.078 nm. Concurrently, Max Haider's team advanced multipole correctors for transmission electron microscopes, influencing designs and allowing routine atomic-resolution imaging of complex structures like defects in oxides. These corrections, commercialized by the mid-2000s, increased signal-to-noise ratios in by factors of up to 10, facilitating quantitative analysis of light elements alongside heavy atoms. From the onward, ADF imaging integrated with (cryo-EM) for beam-sensitive biological samples, combining low-temperature stages with HAADF to preserve native structures while providing mass-thickness contrast. This approach, refined in the , enabled 3D tomography of proteins and cellular components at resolutions of 2-5 nm, as shown in variable-angle HAADF studies of vitreous ice-embedded specimens. Parallel advances in AI-assisted reconstruction, particularly models for denoising, reduced noise in low-dose ADF datasets by 50-70%, improving signal recovery in sparse acquisitions for dynamic biological processes. In the 2020s, low-dose ADF techniques for beam-sensitive materials like metal-organic frameworks achieved atomic-level imaging with doses under 10 electrons/Ų, minimizing damage through and ptychographic enhancements. A significant methodological shift occurred in the with the adoption of segmented and pixelated detectors, replacing analog annular designs to enable simultaneous multi-angle electron collection in 4D-STEM. These detectors, dividing the signal into quadrants or pixels, facilitated differential phase contrast and angular-resolved mapping, capturing variations with sub-milliradian precision and boosting throughput by orders of magnitude for quantitative and field analysis.

Types of Annular Dark-Field Imaging

Conventional ADF

Conventional annular dark-field (ADF) imaging in (STEM) utilizes an annular detector with typical inner collection angles of 10–50 mrad, enabling the capture of electrons scattered at medium angles and rendering it sensitive to both , including thermal diffuse , and to a lesser extent inelastic processes such as plasmons. This configuration distinguishes it from bright-field imaging by excluding the unscattered beam while incorporating a broader range of events compared to stricter high-angle setups. The primary contrast mechanisms in conventional ADF arise from mass-thickness variations across the sample and diffraction effects, leading to an intensity dependence on atomic number (Z) that is approximately Z^{1.5–1.7} but modulated by these factors, making it less exclusively Z-dependent than in high-angle annular dark-field (HAADF) imaging. Diffraction contrast, in particular, introduces coherent contributions from Bragg scattering, which can enhance visibility of structural features but also complicates purely incoherent interpretation. As a result, images often exhibit a mix of and phase-related effects, requiring careful consideration of sample orientation. One key advantage of conventional ADF is its higher signal yield for light elements or thicker samples, where lower-angle scattering provides more electrons to the detector without the rapid attenuation seen at higher angles. This makes it suitable for applications involving beam-sensitive materials or those with lower atomic mass, such as organic nanostructures or light alloys. Furthermore, the inclusion of diffraction effects allows for orientation mapping, where variations in Bragg scattering reveal crystallographic domains and misorientations. A representative example is the of crystalline defects in semiconductors, such as dislocations in , where Bragg scattering from strained lattice regions contributes to localized contrast enhancements, aiding in defect without relying solely on Z-contrast.

Low-Angle ADF (LAADF)

Low-angle annular dark-field (LAADF) imaging is a variation of -STEM that collects electrons scattered at low to medium angles, typically 20–50 mrad inner angle. This setup emphasizes contrast and is particularly useful for visualizing light elements and orientation-dependent features, as it includes more elastically scattered beams compared to higher-angle modes. The contrast is sensitive to and defects, with reduced Z-dependence, making LAADF ideal for mapping fields, phase boundaries, and polycrystalline materials where mass-thickness contrast is minimal. LAADF extends the utility of ADF to beam-sensitive or low-Z samples by providing higher signal levels while highlighting coherent scattering effects. It is often used in combination with other modes for comprehensive analysis, such as in semiconductors or nanomaterials, to reveal features obscured in HAADF due to low scattering efficiency from light atoms.

High-Angle ADF (HAADF)

High-angle annular dark-field (HAADF) imaging is a specialized form of annular dark-field imaging in scanning transmission electron microscopy (STEM) that collects electrons scattered at high angles, typically greater than 50 mrad (or approximately >3°), to minimize contributions from diffracted beams and achieve predominantly incoherent imaging. This configuration ensures that the image intensity arises mainly from thermal diffuse scattering and single elastic scattering events, providing a Z-contrast mechanism where the signal is roughly proportional to the square of the atomic number (Z²) of the scattering atoms. By excluding low-angle scattering associated with Bragg diffraction, HAADF delivers sharp, localized contrast that directly reflects the atomic composition without the phase effects inherent in bright-field or conventional ADF modes. The enhanced sensitivity of HAADF to makes it particularly effective for distinguishing heavy elements within lighter matrices, such as identifying catalytic metal atoms (e.g., or ) dispersed on carbon supports. This Z-dependent contrast allows for the direct of individual heavy atoms or clusters, even at low concentrations, enabling precise compositional at the atomic scale. For instance, in materials like catalysts, HAADF can highlight atoms with high fidelity, as the cross-section scales strongly with , outperforming other modes in . At high collection angles, HAADF exhibits a depth-sectioning effect due to the reduced influence of multiple scattering events, which are more prominent at lower angles and can blur subsurface features. Electrons scattered to these elevated angles primarily result from single, large-angle Rutherford-like interactions, limiting the propagation of along the beam direction and confining the signal to a shallower effective depth within the sample. This property facilitates three-dimensional profiling of atomic distributions, as deeper atoms contribute less to the high-angle signal compared to surface or near-surface ones, aiding in the analysis of layered or buried structures. In aberration-corrected STEM, HAADF is widely employed for direct visualization of atomic structures, bypassing the need for phase reconstruction techniques required in coherent imaging methods like high-resolution TEM. The incoherent nature of HAADF simplifies interpretation, as image intensity monotonically relates to atomic column occupancy and position, allowing sub-angstrom resolution mapping of crystal lattices and defects without artifacts from interference. This capability has become essential for studying complex nanomaterials, where precise atomic-scale insights into structure and chemistry are critical.

Instrumentation and Setup

Detector Geometry

In annular dark-field (ADF) imaging within (), the detector is designed as a ring-shaped annular structure positioned to collect scattered electrons while blocking the unscattered central beam. The central hole in the detector prevents direct beam electrons from reaching the sensor, ensuring that only electrons deflected by the sample contribute to the image signal. This geometry allows for the selection of electrons scattered over a specific annular range, typically defined by inner and outer collection semi-angles, α_i and α_o, respectively. The inner semi-angle α_i marks the boundary of the central hole, excluding low-angle scattering such as the direct beam and small-angle , while the outer semi-angle α_o sets the maximum collection angle, capturing higher-angle scattered electrons. For conventional ADF setups, typical values might include α_i ≈ 5–20 mrad and α_o ≈ 50–100 mrad, whereas high-angle ADF (HAADF) configurations often use larger ranges like α_i ≈ 50 mrad and α_o ≈ 200 mrad to emphasize incoherent for Z-contrast . These angles can be adjusted experimentally; for instance, in variable-angle HAADF, settings of α_i = 60 mrad and α_o = 390 mrad have been achieved at a camera of 100 mm. Detector types vary to meet different needs, with single-ring annular detectors providing basic of scattered over the entire for standard signals. More advanced segmented detectors, such as those with 16 quadrants, or pixellated detectors that divide the annular area into numerous pixels (e.g., for 4D-STEM), enable angular-resolved by isolating signals from specific azimuthal or radial sectors. Segmented and pixellated designs offer enhanced capabilities, such as differential phase contrast or ptychographic reconstruction, by allowing post-processing of spatially variant scattering data, though they require higher data throughput compared to single-ring systems. The detector is positioned in the far-field plane of the column, downstream of the sample, where the diffraction pattern is formed. Effective collection angles are tuned by varying the camera length L, the distance from the sample to the detector plane, using projector lenses; the semi-angle α is approximately given by α ≈ r / L, where r is the radial distance on the detector from the beam center. This adjustment scales the angular acceptance without physically resizing the detector, enabling optimization for different scattering regimes. Calibration of the detector geometry ensures accurate angular definition and minimal overlap with bright-field or patterns, typically achieved by standard samples like undoped to measure beam position and adjust for aberrations or misalignment. For segmented detectors, individual pixel or segment responses are balanced using far-field from a polycrystalline sample to verify uniform collection across the annulus. Such procedures confirm that the inner hole fully blocks the unscattered beam while the outer radius captures the desired angles without contamination from unwanted orders.

Microscope Specifications

Annular dark-field () imaging in () demands precise probe-forming to generate a focused with a spot size below 1 nm, enabling atomic-scale resolution. This is achieved through a series of condenser lenses that demagnify and converge the from the source, typically a , to form the probe. Aberration correctors, such as hexapole or octopole systems (e.g., CEOS hexapole correctors), are integrated into the probe-forming aperture to compensate for spherical and chromatic aberrations, reducing the probe diameter to sub-angstrom levels (e.g., <0.1 nm) while maintaining sufficient current (e.g., >100 pA) for adequate signal intensity. The acceleration voltage for STEM equipped for ADF imaging typically spans 80–300 , selected to optimize the trade-off between and sample integrity. Higher voltages (e.g., 200–300 ) shorten the de Broglie wavelength, enhancing for imaging fine structures, but increase beam penetration and potential in beam-sensitive specimens. Lower voltages (e.g., 80–120 ) reduce damage by limiting energy deposition while still supporting sub-nanometer probes, particularly with aberration correction. Mechanical and environmental stability is essential for sub-angstrom precision in ADF-STEM, necessitating advanced platforms that attenuate low-frequency disturbances (<1 Hz) from building or external sources. Electromagnetic shielding enclosures around the microscope column and laboratory further prevent interference from AC power lines or nearby equipment, minimizing probe drift and ensuring stable scanning over extended periods. Seamless integration with spectrometers for concurrent electron energy-loss spectroscopy (EELS) and energy-dispersive X-ray (EDX) analysis requires coaxial alignment of the probe with detector axes to avoid signal attenuation during ADF imaging. This setup enables simultaneous collection of structural and chemical data, with systems like Gatan Tridiem GIF for EELS and Oxford X-Max detectors for EDX positioned to capture transmitted or scattered electrons without compromising ADF signal fidelity.

Imaging Performance

Resolution Determinants

The spatial resolution in annular dark-field (ADF) imaging is fundamentally governed by the size of the incident electron probe, which sets the minimum achievable detail in the image. The probe diameter d is approximated by the diffraction limit d \approx 0.61 \lambda / \alpha_c, where \lambda is the de Broglie wavelength of the electrons and \alpha_c is the convergence semi-angle of the probe. This limit arises from the wave nature of electrons, with higher accelerating voltages reducing \lambda (e.g., \lambda \approx 0.025 Å at 200 kV) and larger \alpha_c allowing tighter focusing. Aberration correction plays a critical role in approaching this limit by compensating for spherical and chromatic aberrations, enabling probe sizes as small as ~0.5 Å in modern instruments. Without correction, aberrations broaden the probe, degrading resolution to several angstroms; with correction, atomic-scale imaging becomes routine for heavy elements in high-angle ADF modes. The signal-to-noise ratio (SNR) further modulates practical resolution, particularly in beam-sensitive samples where low probe currents or short dwell times are necessary to avoid damage. In ADF imaging, the detected intensity I from scattered electrons obeys Poisson statistics, with SNR scaling as \sqrt{N_\text{scattered}}, where N_\text{scattered} is proportional to the probe current and dwell time per pixel. Higher currents increase N_\text{scattered} and thus SNR, but excessive current can induce specimen drift or damage, limiting effective resolution; conversely, short dwell times (e.g., <1 μs) reduce SNR, blurring fine features unless compensated by averaging multiple frames. Optimizing these parameters—typically balancing currents of 10–100 pA with dwell times of 5–50 μs—ensures that statistical noise does not dominate over the probe size limit. An additional constraint is the delocalization inherent to the scattering process, which smears the signal beyond the probe diameter. For high-angle scattering (\theta > 50 mrad), delocalization is minimal (~0.1–0.5 ) due to small impact parameter b, enabling near-probe resolution for heavy atoms; however, for light atoms, lower Z requires larger b to achieve the same \theta via , increasing delocalization to 1–2 and reducing effective . In the 2020s, techniques have pushed effective resolutions below these hardware constraints by post-processing noisy images. Deep learning-based denoising, such as diffusion models or convolutional neural networks, reconstructs atomic details from low-dose data, achieving sub-Ångström effective resolution even in uncorrected . These methods leverage simulated training data to suppress Poisson noise while preserving structural fidelity, enabling sub-Ångström resolutions in beam-sensitive materials without increasing dose. The probe size itself remains tied to optical performance, as outlined in microscope specifications.

Limitations and Artifacts

One significant limitation of annular dark-field (ADF) imaging in (STEM) is beam damage to sensitive samples. High-angle , which underpins the Z-contrast , requires intense probes to generate adequate signal intensity, exacerbating damage s such as and knock-on displacement in beam-sensitive materials like organics and biological specimens. Organics tolerate approximately 10³ s per square , while biological specimens are limited to ~10 s per square before structural degradation occurs, severely restricting imaging time and resolution for such samples. Various artifacts can also compromise image fidelity in ADF-STEM. Scan distortions, stemming from instabilities in the scanning fields, manifest as wavy patterns and non-orthogonal deformations, particularly noticeable in high-resolution aberration-corrected . In crystalline specimens, channeling effects—where the is steered along columns—induce false variations, displacing apparent positions by several picometers and misleading interpretations of or . Sample thickness imposes another key constraint through multiple scattering effects. As thickness exceeds about 50 nm, repeated elastic and events broaden the angular distribution of scattered electrons, diminishing the high-angle specificity essential for Z-contrast and leading to reduced image and . This limitation is pronounced in substrates supporting nanoparticles, where central fades significantly beyond this thickness threshold. Additionally, imaging exhibits lower dose efficiency compared to bright-field methods, especially for light elements that scatter predominantly at low angles rather than the high angles collected in ADF detectors. This inefficiency necessitates longer acquisition times or higher doses to achieve comparable signal-to-noise ratios, further challenging applications involving low-Z materials and amplifying beam damage risks.

Applications

Materials Characterization

Annular dark-field (ADF) imaging, particularly in the high-angle annular dark-field (HAADF) mode, plays a crucial role in materials characterization by providing Z-contrast sensitivity that highlights atomic number differences, enabling the visualization of heavy elements against lighter matrices in inorganic and composite materials. This technique excels in rigid, high-Z systems such as semiconductors, alloys, and catalysts, where it reveals structural and compositional details at the atomic scale without requiring staining or complex sample preparation. By leveraging incoherent scattering, ADF imaging achieves high signal-to-noise ratios for direct interpretation of atomic positions and distributions. In atomic-scale structure determination, ADF imaging has been instrumental in locating individual dopant atoms within semiconductor lattices, such as heavy impurities like in . For instance, aberration-corrected HAADF-STEM allows direct visualization of single atoms embedded in bulk-like environments, confirming their positions and enabling studies of substitutional incorporation. This capability, often combined with techniques like (EDS), has advanced understanding of distribution in materials like phosphorus-doped delta-layers, where contrasts reveal layer widths and concentrations at sub-nanometer resolution. For interface analysis, ADF imaging visualizes grain boundaries and precipitates in alloys by exploiting Z-contrast to map compositional variations across interfaces. In Ni-rich Ni-Au alloys, HAADF-STEM images delineate bulk interfaces and segregation profiles, showing bright contrasts from heavy Au atoms at boundaries. Similarly, in magnesium alloys, atomic-resolution HAADF reveals co-segregated atoms at grain boundaries, aiding in the study of complexion formation and mechanical properties. This approach facilitates composition mapping of precipitates, such as ordered Cu segregation in boundaries, without the need for additional spectroscopic confirmation in initial structural assessments. In catalyst studies, ADF imaging locates and characterizes metal nanoparticles on oxide supports, critical for optimizing activity and stability. HAADF-STEM images of nanoparticles on CeO₂ supports show their and size distribution, with bright Pt contrasts against the darker CeO₂ matrix, as seen in systems reduced under for oxidation. This visualization has revealed atomic-level fluxional behavior in Pt/CeO₂, where single atoms and clusters migrate during reaction conditions, correlating structure with enhanced catalytic performance. Such imaging guides the design of supported catalysts by quantifying nanoparticle-support interactions. Integration of ADF imaging with electron energy-loss spectroscopy (EELS) enables HAADF-guided elemental quantification at atomic columns, combining structural imaging with chemical analysis. In perovskite-related oxides, HAADF images direct EELS acquisition to specific columns, allowing quantification of coordination numbers and oxidation states via analysis. This tandem approach has quantified elemental ratios in heterostructures, such as Cu-Zn-S interfaces, providing atomic-resolution maps of composition that inform defect chemistry and functionality. By aligning the probe with HAADF-identified features, EELS achieves precise, site-specific quantification with minimal delocalization.

Biological and Nanostructure Analysis

Annular dark-field (ADF) imaging has proven particularly valuable in biological applications due to its sensitivity to mass-thickness contrast, enabling visualization of beam-sensitive samples under low-dose conditions. In cryo-ADF, frozen-hydrated protein samples are imaged to map heavy-atom labels within macromolecules, such as metal ions bound to . This approach detects isolated atoms of and iron with atomic precision, leveraging the incoherent scattering that enhances contrast for high-Z elements in low-Z biological matrices. Recent advancements in low-dose cryo-electron combined with 4D-STEM have achieved sub-nanometer in 3D reconstructions of single protein particles, minimizing while preserving structural details. For nanoscale organic structures, ADF imaging excels in revealing defects and morphologies in carbon-based nanomaterials, capitalizing on its mass-thickness sensitivity to differentiate subtle variations. In carbon nanotubes, low-voltage ADF-STEM provides high-contrast images of multi-walled structures at resolutions better than 2 , highlighting wall defects and alignments without . Similarly, for , aberration-corrected ADF-STEM visualizes atomic-scale defects, such as vacancies and substitutions, by directly imaging local and distortions, which inform electronic property modifications. In soft matter systems, low-angle ADF (LAADF) imaging highlights density variations in organic assemblies, offering contrast for unstained samples where phase contrast alone is insufficient. For polymer blends, ADF-STEM tomography reconstructs 3D morphologies, revealing phase separations and nanoparticle distributions in block copolymers at cryogenic temperatures. In lipid bilayers, LAADF detects thickness fluctuations and domain formations in model membranes, providing insights into biomolecular interactions under hydrated conditions. These techniques are constrained by dose limitations in cryo setups, where electron doses must remain below 10-20 electrons/Ų to avoid structural alterations. Advancements in the 2020s have integrated ADF imaging with for of viral nanostructures, enhancing understanding of and mechanisms. Variable-angle HAADF-STEM improves and reduces artifacts in thick biological sections, enabling precise mapping of viral capsid architectures in samples like SARS-CoV-2. Cryo-electron using ADF detectors has unraveled membrane rearrangements during coronavirus replication, correlating 2D projections with volumetric models at near-atomic . This correlative approach has also facilitated studies of large DNA viruses, where STEM visualizes internal components within intact capsids.

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