Three-dimensional electron diffraction (3D ED) has become increasingly popular over the last two decades, challenging the limits of established single-crystal X-ray diffraction experiments. In particular, continuous-rotation and precession-assisted protocols have been established as important tools in the collection of electron diffraction data, and the data for most of the crystal structures solved by 3D ED nowadays are acquired with one of these two methods. This is particularly true for ceramic materials, where 3D ED allows deep insights into complex structure-property relationships. As ceramic syntheses tend to yield thick and highly crystalline particles, one issue in the refinement against electron diffraction data is the effect of coherent inelastic scattering. While dynamical refinement procedures allow the simulation of multiple scattering events, in general they ignore the inelastic contribution. This work aims to evaluate the impact of dynamical inelastic effects on the diffraction data of ceramics and how their overall quality depends on the measurement strategy used. This was done by comparing the structure models derived from the same crystals under similar experimental conditions of three ceramic compounds, namely NATP [Na1+xAlxTi2-x(PO4)3], LATP [Li2-xAlxTi2-x(PO3)4] and (Fe3Al2[SiO4]3), based on precession, continuous-rotation and stepwise static tilt data. The different methods allowed the detection of low-occupancy sites in the difference electrostatic potential maps corresponding to mobile sodium and lithium ions, paving the way for future studies on their migration behaviour in solid-state electrolytes.

Optimizing the performance of organic solar cells hinges on a comprehensive understanding of their nanostructures, yet traditional characterization methods often fall short, delivering incomplete structural snapshots. We introduce elastically filtered 3D electron diffraction as technique to bridge full reciprocal- and real-space structural analysis within a single transmission electron microscope. Using model bulk heterojunction DRCN5T:PC71BM, 3D electron diffraction reproduces key structural parameters obtained from grazing-incidence wide-angle X-ray scattering, including lattice spacings, coherence lengths, and mosaicity, while also providing true in-plane access and direct registration with high-resolution imaging, diffraction imaging and nano-spectroscopy on the same sample. Application to another archetypal blend, P3HT:PC71BM, demonstrates the generality of the method. Our findings underscore the transformative potential of 3D electron diffraction, particularly in analyzing beam-sensitive organic thin films. The method enables correlative structural characterization of organic solar cells and opens pathways for application to a wide range of other nanostructured materials.

Zeolite Beta, a prototypical intergrowth zeolite with a three-dimensional (3D) system of intersecting 12-member-ring micropore channels, is extensively employed as a versatile solid-acid catalyst. Its structure typically appears as a nanoscale intergrowth of polymorphs arising from the stacking disorder of topologically equivalent layers. Rational regulation of the Beta polymorph composition is, therefore, crucial for optimizing catalytic performance. Here, we report a heterostructure seed-assisted crystallization strategy to synthesize a polymorph E-enriched Beta zeolite (Beta-ER). The 3D electron diffraction, high-resolution transmission electron microscopy imaging, and DIFFaX simulation show that Beta-ER is an intergrown structure of polymorph E/D with polymorph E as the main phase (∼80%). Powder X-ray diffraction, Fourier transform infrared, Raman, and 19F magic-angle spinning nuclear magnetic resonance characterizations suggest that ITQ-1 seeds partially deconstruct in the early stage of crystallization and generate secondary structural building units, which subsequently promote the formation of Beta-ER. In the etherification of FA, Beta-ER with moderate acidity and open pore channels delivers a higher furfuryl ethyl ether (FEE) yield (75.4%) after 30 min at 393 K, outperforming the commercial ZSM-5 (14.2%), Beta (55.9%), and BEC (57.2%) zeolites. Beta-ER maintained stable activity after four regeneration cycles. Density functional theory calculations confirm that substituting framework Al with Ge increases the energy barrier for the ring opening of FEE, thereby suppressing the formation of byproducts. In addition, kinetic analysis and molecular dynamics simulations reveal that the interconnected channel system of Beta-ER facilitates the diffusion of substrates and products.

The crystalline sponge (CS) method utilizes a crystalline porous material to arrange target molecules within its periodic pores. This enables the determination of the 3D atomic structures of organic molecules without the need for crystallization. However, its applicability is currently limited by the availability of suitable porous single crystals that can grow to a sufficient size for X-ray diffraction analysis. Although three-dimensional electron diffraction (3D ED) allows structure determination from nanosized crystals, ab initio structural analysis of organic molecules hosted in nanocrystalline sponges remains challenging and largely manual. Here, we present a 3D ED-based nanocrystalline sponge (NanoCS) workflow that integrates guest soaking, low-dose cryogenic data collection, and automated structure solution and refinement. A key advance is a newly developed automated approach for guest identification and structural analysis implemented in the AutoSolveX pipeline. Using the nanocrystalline bismuth-based metal-organic framework (MOF) SU-100 as a prototype crystalline sponge, we demonstrated the general applicability of this NanoCS strategy. 10 organic molecules, introduced as pure liquids, solutions, or vapors, are investigated. For all systems, 3D ED data collected under low electron fluence and cryogenic conditions enabled fully automated identification and refinement of the guest molecules using AutoSolveX. The results confirm the periodic arrangement of the guest molecules within the pores of SU-100, mediated by coordination bonding, hydrogen bonding, offset π-π stacking, and van der Waals interactions. This work establishes NanoCS combined with automated structural analysis as a practical and high-throughput platform for routine ab initio structural determination of organic molecules from nanocrystalline hosts.

Diamine functionalization of the metal-organic framework Mg2(dobpdc) (dobpdc4- = 4,4'-dioxidobiphenyl-3,3'-dicarboxylate) significantly enhances its selectivity for CO2 capture from flue gases and air. The structure and CO2 capacity of such materials are typically assessed using bulk techniques that rely on averaging signal over large ensembles of unit cells, obscuring local heterogeneities, such as variations in CO2 occupancy across individual nanocrystals. To resolve this limitation, we demonstrate a multimodal, nanoscale characterization of Mg2(dobpdc) appended with 1,3-diaminopropane. By employing recently developed characterization techniques at progressively smaller length scales, we uncover insights from correspondingly smaller populations of unit cells. First, we use parallel-beam 3D electron diffraction (3D ED) to identify a prominent expansion in lattice parameters upon desorption of CO2, as observed at the level of single nanocrystals. Second, we use convergent-probe 4D scanning transmission electron microscopy (4D-STEM) to quantify associated differences in lattice strain as a function of gas loading and diamine appending. These measurements sample small subvolumes within individual nanocrystals. Finally, we apply infrared scattering scanning near-field optical microscopy (IR s-SNOM) to confirm variable CO2 chemisorption across adsorption sites at the surface of single nanocrystals. This multimodal, multiscale approach allows us to map heterogeneity within individual nanocrystals. Collectively, these findings emphasize the importance of local, nanoscale characterization of metal-organic frameworks in revealing previously unresolvable features that impact their performance.

Abstract To overcome the limitations of traditional single-crystal X-ray diffraction (SCXRD) for microcrystalline materials and the peak-overlapping issue of powder X-ray diffraction (PXRD), this study employed cryogenic continuous rotation electron diffraction (cryo-cRED) with a low-dose strategy to determine the crystal structure of CL30, a novel silicogermanate framework. It is confirmed that CL30 crystallizes in the C 2/ m space group and has layered topology composed of discontinuous zigzag chains connected by double four-membered ring (d4r) units, with fluoride anions (F − ) occluded in the d4r units. In CL30, charge balance involves organic structure-directing agent (OSDA) cations, occluded F − , and terminal oxygen sites whose protonation state cannot be established from the present three dimensional (3D) ED data. F − encapsulated in the d4r units contributes to charge compensation as the counter-anion to OSDA cations, rather than only balancing the framework charge. Although the refinement indices ( R 1 = 0.29, wR 2 = 0.71) exceeded typical small-molecule crystallography standards, the structural model remained highly reliable, as supported by geometric restraints and validation. In electron diffraction, elevated R 1 values are commonly attributed to the intrinsic factors of the technique, such as dynamic scattering, detector noise from scintillator-based detectors, and TEM stage instability (large spheres of confusion). This study introduces a new structural prototype to the silicogermanate family and establishes a feasible workflow for determining the structures of radiation-sensitive microcrystalline porous materials.

Intracellular crystallization is an emerging approach in structural biology that bypasses the need for protein purification. In 2024, the InCellCryst pipeline was introduced for structural studies of intracellular crystals by serial X-ray crystallography. Serial crystallography requires the exposure of tens of thousands of cells containing intracellular crystals, precluding high-resolution studies on proteins that crystallize only in a few cells. Here we introduce IncelluloED, a method that combines intracellular crystallization with in situ 3D electron diffraction in cells and achieves high-resolution structures from just one crystal inside one cell. Experiments on a microcrystal of the HEX-1 protein from Magnaporthe grisea, grown inside an insect cell, give a structure at 1.9 Å resolution from a volume of ~1.6 µm3 as compared to 1.8 Å resolution achieved by serial X-ray crystallography from a combined volume exceeding eleven million µm3. IncelluloED uses widely available cryo-EM tools and brings high-resolution structural biology into home laboratories while also advancing a vision for a "single-cell structural laboratory".

The crystal structure of Rb2Ru2O7·H2O was determined by three-dimensional electron diffraction from the individual crystallites of a solid-state powder product. Rb2Ru2O7·H2O crystallizes in space group C2/c (a = 7.841(3) Å, b = 12.500(3) Å, c = 8.392(2) Å, β = 93.57(4)°, Z = 4). The structure contains infinite chains that run normal to the (101) plane and consist of alternating RuO6 octahedra and square-pyramidal RuO5 units connected via shared O-O edges. Magnetic properties were measured on the bulk powder, showing a diamagnetic baseline from 300 K to 60 K with a small Curie tail below 55 K. The magnetic moment, calculated from the 1.8 K isotherm, saturates at M = 4.4 × 10-3 μB Ru-1, much less than would be expected for S = 1 ruthenium. Bond-valence-sum analysis indicates high-valent Ru, and the near-diamagnetic response is consistent with the edge-sharing Ru-Ru motif, where weak direct Ru-Ru overlap yields a local singlet.

2D covalent organic frameworks (COFs) usually possess a polycrystalline nature as well as lower porosity and surface area than 3D counterparts, restraining their exploration over gas storage applications. Herein, a substituent strategy has been proposed and employed to generate three robust single-crystal 2D COFs isomers with atom-resolution structures determined by 3D electron diffraction. Among three isomers, a precise engineering of their interlayer distance affords the highest Brunauer-Emmett-Teller surface area of ~2100 m2 g-1 and the largest pore volume of 1.40 cm3 g-1 for the desolvated GZU-1. This COF shows the highest total volumetric methane uptake of 240 cm3 (STP) cm-3 at 273 K and 100 bar among 2D COFs, even comparable with those for excellent 3D MOFs. This work not only delivers unique insight into the design of 2D single-crystal COFs by interlayer stacking regulation, but also promotes the application of highly porous 2D COFs in gas storage.

3D electron diffraction (3D ED) has undergone impressive development in the last decade. However, its accuracy and reproducibility have never been tested, up to now, in different laboratories on the same batch of samples. This paper reports a round robin on three test structures, two inorganic and one organic, solved and refined with 3D ED in seven different laboratories employing different transmission electron microscopes, with different acceleration voltages, different methodologies and different detectors. The results of the round robin show a remarkable accuracy of the technique that, in the case of kinematical refinement, is around 0.05 Å error on atomic positions for the inorganic samples and 0.15 Å for the beam-sensitive organic crystal. Dynamical refinement further improves the accuracy. The analysis of diverse samples and numerous data sets again confirms that dynamical refinement is a well established procedure, significantly reducing the refinement R factors, improving the accuracy of the structure models in most cases, and providing fine structural details, such as hydrogen-atom positions and the absolute structure, for both inorganic and organic samples.

The crystallization of a trimetallic cobalt-molybdenum-sodium metal-organic <!?tlsb=-0.05pt>framework, poly[μ-benzene-1,3,5-tricarboxylato-tetra-μ-oxido-cobaltmolybdenumtrisodium], UOW-10 or [Na3Co(MoO4)(BTC)]n, is achieved by solvothermal synthesis using benzene-1,3,5-tricarboxylic acid (H3BTC, C9H6O6) as a ligand precursor, Na2MoO4·2H2O and Co(NO3)2·2H2O as metal sources, and N,N-dimethylformamide (DMF) as the solvent. 3D electron diffraction (3D ED) reveals that the structure crystallizes in the monoclinic space group P21/c, with lattice parameters of a= 9.718 (2), b= 18.250 (3), c= 6.892 (9) Å, α= γ= 90, β= 96.156 (15)°, V= 1214.7 (4) Å3 and Z= 4. The phase purity of the bulk sample was confirmed using synchrotron powder X-ray diffraction. The organic ligands form a 2D layer, where cobalt and molybdenum are found, with sodium cations located between the layers. There are four crystallographically distinct sodium sites: three exhibit a distorted octahedral coordination geometry, while the remaining site is seven-coordinate. The cobalt has trigonal bipyramidal coordination geometry and molybdenum exhibits a tetrahedral coordination geometry. Half the sodium cations in the structure forms 1D column-like motifs via shared oxygen edges along the crystallographic c axis, which are cross-linked in b by the cobalt and molybdenum sites via bridging O atoms, while the other half of the sodium cations form 2D ribbons in the ac plane, propagating along c, linked by sharing oxygen edges and faces. The optical properties of UOW-10 were investigated through the use of UV-Vis spectroscopy, showing a bandgap of 1.8 eV. Deconvolution of the features in the visible-light region reveals that four peaks are present, which can all be ascribed to the d-d transitions from the trigonal bipyramidal cobalt. By means of thermogravimetric analysis (TGA) and variable-temperature powder X-ray diffraction (VT-PXRD), it is demonstrated that the material has thermal stability to 410 °C, after which structure collapse occurs, leading to a mixture of Na2MoO4, CoO and Co3Mo above 900 °C.

While 3D electron diffraction (3D-ED or microcrystal electron diffraction; MicroED) has emerged as a promising method for protein structure determination, its applicability is hindered by a high susceptibility to radiation damage, leading to a decreasing signal-to-noise ratio in consecutive diffraction patterns that limits the quality (resolution and redundancy) of the data. In addition, data completeness may be restricted due to the geometrical limitations of current sample holders and stages. Although specialized equipment can overcome these challenges, many laboratories do not have access to such instrumentation. In this work, we introduce an approach that addresses these issues using a commonly available 200 keV cryo-electron microscope. The multi-position acquisition technique that we present here combines (i) multiple data acquisitions from a single crystal over several tilt ranges and (ii) merging data from a small number of crystals each tilted about a different axis. The robustness of this approach is demonstrated by the de novo elucidation of a protein-peptide complex structure from only two orthorhombic microcrystals.

Spin crossover (SCO) materials hold exceptional potential for applications in sensors, memory and photonic devices, and actuators, but their practical use remains constrained by a scarcity of compounds exhibiting transition temperatures above room temperature. Herein, we report a postsynthetic strategy to incorporate silver atoms within the pores of a Hofmann-type clathrate {Fe(pyrazine)[Ni(CN)4]} framework. This in situ silver integration induces an 80 K shift in the spin transition temperature, accompanied by a 45 K wide hysteresis loop. Structural analyses using high-resolution TEM, PXRD, 3D electron diffraction, and spectroscopic techniques reveal that silver insertion proceeds via a redox-driven Ni(II) to Ni(III) transformation, leading to partial decoordination of the framework and the formation of silver atoms.

Cordyceps species are widespread entomopathogens and promising biocontrol agents that produce diverse secondary metabolites, yet the roles of these molecules during the infection process remain unclear. To interpret how fungal chemistry contributes to host colonization, we compared the metabolomes and virulence traits of two strains of phylogenetically distinct Cordyceps species (C.javanica and C.blackwelliae) and assessed their effects on beet armyworms (fungiSpodopteraexigua). Virulence assays revealed species-dependent pathogenicity, with C.javanica showing the highest virulence. Combining untargeted metabolomics, feature-based molecular networking (FBMN), 3D electron-diffraction crystallography and comprehensive 1D/2D NMR, we gained insights into their metabolomic traits. For instance, C.javanica displayed notable beauveriolide diversity, including three previously undescribed derivatives (1–3), while C.blackwelliae produced mainly diketopiperazines in vitro. The FBMN results revealed putative beauveriolide analogs in the C.blackwelliae extracts, unlike the cadaver analysis, revealing beauvericins in infected corpses. Remarkably, the crude extracts obtained from authentic insect cadavers contained beauveriolides and beauvericins, providing in vivo chemical evidences of their production during infection for the first time. Moreover, bioassays with purified compounds showed that insecticidal activity cannot be attributed across all beauveriolides but depends on amino-acid composition, implying multifunctional roles beyond direct toxicity. Altogether, these results reveal context-dependent metabolic reprogramming and species-specific chemical strategies in entomopathogenic fungi, with implications for microbial ecology, host specificity, and the rational development of fungal biocontrol agents. The results of this study also give rise to the need for more intensified study on the chemical composition of the insect cadavers that are colonized by other entomopathogens.

Determining chirality in crystalline powders through 3D electron diffraction.

The MIL‐53 series of metal–organic frameworks (MOFs) is considered archetypal for flexible MOFs, with desolvated materials rapidly transitioning between open and closed phases in a stepwise breathing process in response to changes in temperature under ambient pressure conditions. Herein, the differing structures of MIL‐53(Cr) and MIL‐53(Ga) during breathing – hydrated and anhydrous, closed and open pore–are characterized by in situ single crystal 3D electron diffractionthrough varying sample conditions within the electron diffractometer. In doing so, the crystal structures of nine intermediate phases are uncovered that together represent the continuous breathing of MIL‐53 under vacuum, in stark contrast to ambient pressure stepwise breathing. In addition, these structures offer insight into particle‐to‐particle structural heterogeneity that are averaged out by conventional powder X‐ray diffraction measurements, and may begin to explain metal‐dependent adsorption phenomena observed across the MIL‐53 homologues. In situ 3D electron diffraction is therefore expected to become a powerful tool for in‐depth structural investigations of flexible porous materials.

Cyanido (CN−)‐bridged coordination polymers (CP) have been extensively studied as molecular‐based functional materials. However, synthesizing 3D compounds composed only of metal ions and CN−—without bulky organic groups—and that melt before decomposing remains a considerable challenge. This difficulty arises because CN− strongly interconnect metal ions, forming rigid, dense frameworks with high melting points. In this study, we successfully synthesized a melting composite consisting of 3D KCd[Cu(CN)2]3 and 2D K2Cu3(CN)5 by dehydrating K2Cd(H2O)Cu4(CN)8·1.5H2O. Remarkably, nanodomains of these two compounds coexisted within single particles, allowing their crystal structures to be independently determined by 3D electron diffraction (MicroED) of the resulting powders. Each compound melted at its respective melting point, around 559 K. Notably, the melting point of KCd[Cu(CN)2]3 is unusually low for a 3D dense coordination framework. This atypically low melting point results from a combination of crystalline surface effects, and the entropy contribution of the dynamic, labile two‐coordinate Cu centers in the framework. Additionally, we demonstrated a reversible transformation between the dehydrated mixture and the hydrated parent compound through exposure to water vapor, highlighting the dynamic and responsive nature of these CN−‐based solid‐state materials.

Three-dimensional electron diffraction (3D ED) has emerged as a powerful tool for solving the structures of small crystals down to nanometre-scale sizes. Despite advancements in automating data acquisition for 3D ED, the subsequent data processing and structure solution have largely relied on human intervention and have been mostly conducted offline. This reliance on expertise in electron crystallography and the lack of real-time feedback on data quality and structural information have limited the broader adoption of 3D ED. Here, we introduce Instamatic-solve, a fully automated, real-time structure solution pipeline for 3D ED deployed on a JEOL JEM 2100 transmission electron microscope. Instamatic-solve streamlines the entire process by automating the subsequent data processing and structure solution, providing real-time assessments of data quality and structural information. Moreover, the pipeline can handle offline 3D ED data acquired from various transmission electron microscope platforms. Using Instamatic-solve, we have successfully solved the crystal structures of diverse materials, including seven inorganic zeolites, two inorganic-organic hybrids and four organic molecules (including pharmaceuticals), all within 2 min. Instamatic-solve mimics the typical manual structure solution process, and its outcomes depend heavily on data quality. Our results indicate that a routine and reliable structure solution is achievable in most cases, provided that the data meet critical quality criteria, namely completeness ≥50% and resolution better than 1.0 Å. By enabling efficient, automated and real-time structure solution for crystalline materials, Instamatic-solve spans various scientific disciplines.

Many semiconductortechnologies require interfacing materials withdifferent properties. 2D hybrid perovskites are one of the most promisingcandidates, combining the advantages of organic and inorganic layers.The networks of linked metal-halide octahedra with voids filled byorganic counterions have proven high variability and can be tailoredto specific applications. The geometric and electronic setup of theorganic linker molecule between inorganic layers affects the crystalstructure and the overall optoelectronic properties. Monoamines typicallyform bilayers in so-called Ruddlesden–Popper phases (RPs),while bisamines allow for making Dion–Jacobson phases (DJs),with only a monolayer directly bridging the inorganic layers. Therefore,it would be highly interesting if one could compare RPs to DJs directlyto each other, meaning that they have been prepared using exactlythe same organic linker molecule, which is the aim of the study presentedhere. Because of the potential interaction of π-conjugated compoundswith the electronic system of the semiconductor, we have selecteda special linker here: a divalent ferrocene derivative containingone primary amine attached to each of the cyclopentadienyl rings.These linkers form novel quasi-DJs, and their structure was determinedby 3D electron diffraction and density functional theory. We foundthat by different crystallization kinetics, two quasi-DJ variantsand even RPs can be obtained from the same spacer molecule. It takestime for the ferrocene-based linker to adjust to a particular conformation,giving the system also time to form octahedral connections other thanthe classic DJ/RP corner-sharing. The different octahedral linkages,ranging from face- to corner-sharing, have a significant influenceon the optoelectronic properties. Thus, with a change of the synthesisconditions, it is possible to change the structure and the propertiesof the same educts. Our study also enables the first direct comparisonof quasi-DJ and RP phases by achieving both with the same spacer molecule.

The topological insulator Bi14Rh3I9, a layered salt, is exfoliated through a heterogeneous reaction with n‐butyllithium and subsequent sonication. Precession‐assisted 3D electron diffraction tomography of a nanodomain of an exfoliated platelet revealed that the material is reduced to the intermetallic superconductor Bi14Rh3. The orthorhombic structure of Bi14Rh3 is a 3D polyhedra framework of edge‐sharing [RhBi8/2] cubes and square antiprisms that has isolated Bi– anions in its helical channels. The kagome‐type layer Bi12Rh3]3+ of the precursor Bi14Rh3I9 and the Bi12Rh3]2+ framework of the product Bi14Rh3 have the same composition, very similar structural motifs and interatomic distances, but differ in the dimensionality of the network. The chemical exfoliation, which is based on a redox leaching reaction, is thus accompanied by a topochemical transformation that involves a massive atomic rearrangement in the intermetallic part of the structure. This unconventional and complex process yields an intermetallic compound that exhibits a 3D crystal structure while manifesting the morphology characteristic of a 2D material.