X-ray Diffraction Data Collection Research Articles

Zinc hydroxide sulphate and its transformation to crystalline zinc oxide

The thermal transformation of zinc hydroxide sulphate hydrate to zinc oxide has been examined using synchrotron X-ray diffraction, thermogravimetric analysis, scanning electron microscopy, and surface area measurements. By collecting X-ray diffraction data in situ, we found that the dehydration of zinc hydroxide sulphate pentahydrate proceeded in discrete steps to form anhydrous zinc hydroxide sulphate. This compound then decomposed to a mixture of zinc oxide and a compound tentatively identified as Zn3(OH)2(SO4)2 at ~235 °C. At ~360 °C, the final dehydroxylation occurred with the formation of zinc oxy-sulphate, Zn3O(SO4)2, which then decomposed to ZnO at about ~800 °C. Interruption of the dehydration process can be used to synthesize the intermediate compounds.

A microfluidic approach for protein structure determination at room temperature via on-chip anomalous diffraction

We report a microfluidic approach for de novo protein structure determination via crystallization screening and optimization, as well as on-chip X-ray diffraction data collection. The structure of phosphonoacetate hydrolase (PhnA) has been solved to 2.11 Åvia on-chip collection of anomalous data that has an order of magnitude lower mosaicity than what is typical for traditional structure determination methods.

The Unique Structure of Haemophilus influenzae Protein E Reveals Multiple Binding Sites for Host Factors

Haemophilus influenzae protein E (PE) is a multifunctional adhesin involved in direct interactions with lung epithelial cells and host proteins, including plasminogen and the extracellular matrix proteins vitronectin and laminin. We recently crystallized PE and successfully collected X-ray diffraction data at 1.8 Å. Here, we solved the structure of a recombinant version of PE and analyzed different functional regions. It is a dimer in solution and in the asymmetric unit of the crystals. The dimer has a structure that resembles a flattened β-barrel. It is, however, not a true β-barrel, as there are differences in both the hydrogen-bonding pattern and the shape. Each monomer consisted of a 6-stranded antiparallel β-sheet with a rigid α-helix at the C terminus tethered to the concave side of the sheet by a disulfide bridge. The laminin/plasminogen binding region (residues 41 to 68) is exposed, while the vitronectin binding region (residues 84 to 108) is partially accessible in the dimer. The dimerized PE explains the simultaneous interaction with laminin and vitronectin. In addition, we found this unique adhesin to be present in many bacterial genera of the family Pasteurellaceae and also orthologues in other, unrelated species (Enterobacter cloacae and Listeria monocytogenes). Peptides corresponding to the surface-exposed regions PE 24 to 37, PE 74 to 89, and PE 134 to 156 were immunogenic in the mouse. Importantly, these peptide-based antibodies also recognized PE at the bacterial surface. Taken together, our detailed structure of PE explains how this important virulence factor of H. influenzae simultaneously interacts with host vitronectin, laminin, or plasminogen, promoting bacterial pathogenesis.

Nearest-cell: a fast and easy tool for locating crystal matches in the PDB.

When embarking upon X-ray diffraction data collection from a potentially novel macromolecular crystal form, it can be useful to ascertain whether the measured data reflect a crystal form that is already recorded in the Protein Data Bank and, if so, whether it is part of a large family of related structures. Providing such information to crystallographers conveniently and quickly, as soon as the first images have been recorded and the unit cell characterized at an X-ray beamline, has the potential to save time and effort as well as pointing to possible search models for molecular replacement. Given an input unit cell, and optionally a space group, Nearest-cell rapidly scans the Protein Data Bank and retrieves near-matches.

Room-temperature X-ray diffraction studies of cisplatin and carboplatin binding to His15 of HEWL after prolonged chemical exposure

The anticancer complexes cisplatin and carboplatin are known to bind to both the Nδ and the Nℇ atoms of His15 of hen egg-white lysozyme (HEWL) in the presence of dimethyl sulfoxide (DMSO). However, neither binds in aqueous media after 4 d of crystallization and crystal growth, suggesting that DMSO facilitates cisplatin/carboplatin binding to the N atoms of His15 by an unknown mechanism. Crystals of HEWL cocrystallized with cisplatin in both aqueous and DMSO media, of HEWL cocrystallized with carboplatin in DMSO medium and of HEWL cocrystallized with cisplatin and N-acetylglucosamine (NAG) in DMSO medium were stored for between seven and 15 months. X-ray diffraction studies of these crystals were carried out on a Bruker APEX II home-source diffractometer at room temperature. Room-temperature X-ray diffraction data collection removed the need for cryoprotectants to be used, ruling out any effect that the cryoprotectants might have had on binding to the protein. Both cisplatin and carboplatin still bind to both the Nδ and Nℇ atoms of His15 in DMSO media as expected, but more detail for the cyclobutanedicarboxylate (CBDC) moiety of carboplatin was observed at the Nℇ binding site. However, two molecules of cisplatin were now observed to be bound to His15 in aqueous conditions. The platinum peak positions were identified using anomalous difference electron-density maps as a cross-check with Fo-Fc OMIT electron-density maps. The occupancies of each binding site were calculated using SHELXTL. These results show that over time cisplatin binds to both N atoms of His15 of HEWL in aqueous media, whereas this binding is speeded up in the presence of DMSO. The implication of cisplatin binding to proteins after a prolonged period of time is an important consideration for the length of treatment in patients who are given cisplatin.

Harvesting and Cryo-cooling Crystals of Membrane Proteins Grown in Lipidic Mesophases for Structure Determination by Macromolecular Crystallography

An important route to understanding how proteins function at a mechanistic level is to have the structure of the target protein available, ideally at atomic resolution. Presently, there is only one way to capture such information as applied to integral membrane proteins (Figure 1), and the complexes they form, and that method is macromolecular X-ray crystallography (MX). To do MX diffraction quality crystals are needed which, in the case of membrane proteins, do not form readily. A method for crystallizing membrane proteins that involves the use of lipidic mesophases, specifically the cubic and sponge phases1-5, has gained considerable attention of late due to the successes it has had in the G protein-coupled receptor field6-21 (www.mpdb.tcd.ie). However, the method, henceforth referred to as the in meso or lipidic cubic phase method, comes with its own technical challenges. These arise, in part, due to the generally viscous and sticky nature of the lipidic mesophase in which the crystals, which are often micro-crystals, grow. Manipulating crystals becomes difficult as a result and particularly so during harvesting22,23. Problems arise too at the step that precedes harvesting which requires that the glass sandwich plates in which the crystals grow (Figure 2)24,25 are opened to expose the mesophase bolus, and the crystals therein, for harvesting, cryo-cooling and eventual X-ray diffraction data collection. The cubic and sponge mesophase variants (Figure 3) from which crystals must be harvested have profoundly different rheologies4,26. The cubic phase is viscous and sticky akin to a thick toothpaste. By contrast, the sponge phase is more fluid with a distinct tendency to flow. Accordingly, different approaches for opening crystallization wells containing crystals growing in the cubic and the sponge phase are called for as indeed different methods are required for harvesting crystals from the two mesophase types. Protocols for doing just that have been refined and implemented in the Membrane Structural and Functional Biology (MS&FB) Group, and are described in detail in this JoVE article (Figure 4). Examples are given of situations where crystals are successfully harvested and cryo-cooled. We also provide examples of cases where problems arise that lead to the irretrievable loss of crystals and describe how these problems can be avoided. In this article the Viewer is provided with step-by-step instructions for opening glass sandwich crystallization wells, for harvesting and for cryo-cooling crystals of membrane proteins growing in cubic and in sponge phases.

Harvesting and Cryo-cooling Crystals of Membrane Proteins Grown in Lipidic Mesophases for Structure Determination by Macromolecular Crystallography

An important route to understanding how proteins function at a mechanistic level is to have the structure of the target protein available, ideally at atomic resolution. Presently, there is only one way to capture such information as applied to integral membrane proteins (Figure 1), and the complexes they form, and that method is macromolecular X-ray crystallography (MX). To do MX diffraction quality crystals are needed which, in the case of membrane proteins, do not form readily. A method for crystallizing membrane proteins that involves the use of lipidic mesophases, specifically the cubic and sponge phases1-5, has gained considerable attention of late due to the successes it has had in the G protein-coupled receptor field6-21 (www.mpdb.tcd.ie). However, the method, henceforth referred to as the in meso or lipidic cubic phase method, comes with its own technical challenges. These arise, in part, due to the generally viscous and sticky nature of the lipidic mesophase in which the crystals, which are often micro-crystals, grow. Manipulating crystals becomes difficult as a result and particularly so during harvesting22,23. Problems arise too at the step that precedes harvesting which requires that the glass sandwich plates in which the crystals grow (Figure 2)24,25 are opened to expose the mesophase bolus, and the crystals therein, for harvesting, cryo-cooling and eventual X-ray diffraction data collection.The cubic and sponge mesophase variants (Figure 3) from which crystals must be harvested have profoundly different rheologies4,26. The cubic phase is viscous and sticky akin to a thick toothpaste. By contrast, the sponge phase is more fluid with a distinct tendency to flow. Accordingly, different approaches for opening crystallization wells containing crystals growing in the cubic and the sponge phase are called for as indeed different methods are required for harvesting crystals from the two mesophase types. Protocols for doing just that have been refined and implemented in the Membrane Structural and Functional Biology (MS&FB) Group, and are described in detail in this JoVE article (Figure 4). Examples are given of situations where crystals are successfully harvested and cryo-cooled. We also provide examples of cases where problems arise that lead to the irretrievable loss of crystals and describe how these problems can be avoided. In this article the Viewer is provided with step-by-step instructions for opening glass sandwich crystallization wells, for harvesting and for cryo-cooling crystals of membrane proteins growing in cubic and in sponge phases.

Adenine Binding Mode Is a Key Factor in Triggering the Early Release of NADH in Coenzyme A-dependent Methylmalonate Semialdehyde Dehydrogenase

Structural dynamics associated with cofactor binding have been shown to play key roles in the catalytic mechanism of hydrolytic NAD(P)-dependent aldehyde dehydrogenases (ALDH). By contrast, no information is available for their CoA-dependent counterparts. We present here the first crystal structure of a CoA-dependent ALDH. The structure of the methylmalonate semialdehyde dehydrogenase (MSDH) from Bacillus subtilis in binary complex with NAD(+) shows that, in contrast to what is observed for hydrolytic ALDHs, the nicotinamide ring is well defined in the electron density due to direct and H(2)O-mediated hydrogen bonds with the carboxamide. The structure also reveals that a conformational isomerization of the NMNH is possible in MSDH, as shown for hydrolytic ALDHs. Finally, the adenine ring is substantially more solvent-exposed, a result that could be explained by the presence of a Val residue at position 229 in helix α(F) that reduces the depth of the binding pocket and the absence of Gly-225 at the N-terminal end of helix α(F). Substitution of glycine for Val-229 and/or insertion of a glycine residue at position 225 resulted in a significant decrease of the rate constant associated with the dissociation of NADH from the NADH/thioacylenzyme complex, thus demonstrating that the weaker stabilization of the adenine ring is a key factor in triggering the early NADH release in the MSDH-catalyzed reaction. This study provides for the first time structural insights into the mechanism whereby the cofactor binding mode is responsible at least in part for the different kinetic behaviors of the hydrolytic and CoA-dependent ALDHs.

Will Kleber, Hans-Joachim Bautsch, Joachim Bohm, Einführung in die Kristallographie

This book is the classic German language textbook on Crystallography! It has first been published in 1956. Since then, the (VEB) Verlag Technik, Berlin, published a total of 18 editions up to 1998. In 2010, the Oldenbourg Verlag Munich published the 19th edition, with which this book review is mainly concerned. Up to 1970, i.e., the 10th German edition and 1st English edition (under the title An Introduction to Crystallography), Will Kleber was the sole author. Since then Irmgard Kleber, Hans-Joachim Bautsch, Joachim Bohm, and most recently Detlev Klimm worked on the subsequent editions. All of these authors followed the didactic approach of Will Kleber while incorporating newer results and expanding the references and index. All of crystallography is treated comprehensively under the five main sections: (i) crystal structure and morphology, (ii) crystal chemistry, (iii) physical-chemical crystallography, (iv) crystal physics, and (v) structure analysis of crystals. There are a total of 413 pages of text and an addendum (Wulff net, Schmidt net and birefringence/interference table). Very notable additions of recent years in the crystal physics section are the symmetry groups of fields and Curie's symmetry principle as well as an introduction to the tensor formalism. Otherwise, this reviewer feels that especially the sections on structure analysis and crystal chemistry need updating. With respect to the described crystal structure types, the crystal chemistry section is partially outdated. There are no descriptions of modern materials, modular structures, nano-structured materials, and biological macromolecule structures. This reviewer missed also an introduction to group-subgroup symmetry relations between crystal structures (i.e. Bärnighausen trees). Open-access Internet resources such as the “Bilbao Crystallographic Server” (http://www.cryst.ehu.es/) and the “Crystallography Open Database” (www.crystallography.net) should be mentioned in future editions of the book, since the latter database possesses more than 200,000 (unit cell parameters, space group and setting, atomic coordinates, …, full bibliography) entries. In the structure analysis section, the Weissenberg method (of 1924) and other film-based methods of X-ray diffraction data collection are covered extensively while position sensitive X-ray detectors are not even mentioned. Similarly, transmission electron microscopy (TEM) and electron diffraction are covered within nine pages only that seem to have not changed much over recent years. As a result, instrumental (such as TEM-lens aberration correction) and methodical (e.g., precession electron diffraction based crystallography) breakthroughs are ignored although there is an atomic-resolution phase-contrast TEM image on the front cover of the book. As for this front cover image, it would be of higher pedagogical value if it were to show a length scale marker. Also a model of the crystal structure could be shown on the book cover in three dimensions next to this image (rather than sketched into the two-dimensional experimental data). As far as this reviewer is aware, there were no more English language editions since 1970. This is a real pity as generations of materials science and crystallography students in the English speaking parts of the world have missed out on this classical textbook. There are indeed a few classical books, e.g., John von Neumann’s, Mathematische Grundlagen der Quantummechanik (Julius Springer-Verlag, Berlin, 1932, Springer 1968 and 1996; translated into English 1949: Mathematical Foundations of Quantum Mechanics, Princeton Landmarks in Mathematics, 12th printing, 1996), that never need updating. Will Kleber's book, however, needs updating in future editions because it aims to cover classical and topical theories as well as experiments comprehensively. Because its original approach to introducing crystallography has still lots of merits, a 20th German/2nd English edition of his book would be very valuable for the education of new generations of materials scientists and crystallographers worldwide. Active research scientists/educators such as Detlef Klimm who “grew up” with “The Kleber”, utilized it as main textbook while studying crystallography as undergraduates and/or graduates (and are now in their forties to mid-fifties), could be the authors of such new editions.

Single-drop optimization of protein crystallization.

A completely new crystal-growth device has been developed that permits charting a course across the phase diagram to produce crystalline samples optimized for diffraction experiments. The utility of the device is demonstrated for the production of crystals for the traditional X-ray diffraction data-collection experiment, of microcrystals optimal for data-collection experiments at a modern microbeam insertion-device synchrotron beamline and of nanocrystals required for data collection on an X-ray laser beamline.

Substrate binds in the S1 site of the F253A mutant of LeuT, a neurotransmitter sodium symporter homologue

LeuT serves as the model protein for understanding the relationships between structure, mechanism and pharmacology in neurotransmitter sodium symporters (NSSs). At the present time, however, there is a vigorous debate over whether there is a single high-affinity substrate site (S1) located at the original, crystallographically determined substrate site or whether there are two high-affinity substrates sites, one at the primary or S1 site and the other at a second site (S2) located at the base of the extracellular vestibule. In an effort to address the controversy over the number of high-affinity substrate sites in LeuT, one group studied the F253A mutant of LeuT and asserted that in this mutant substrate binds exclusively to the S2 site and that 1 mM clomipramine entirely ablates substrate binding to the S2 site. Here we study the binding of substrate to the F253A mutant of LeuT using ligand binding and X-ray crystallographic methods. Both experimental methods unambiguously show that substrate binds to the S1 site of the F253A mutant and that binding is retained in the presence of 1 mM clomipramine. These studies, in combination with previous work, are consistent with a mechanism for LeuT that involves a single high-affinity substrate binding site.

X-ray diffraction characterization of polymer intercalated graphite oxide

Graphite oxide (GO) is generated by treating graphite with strong oxidizers. GO retains the structure of graphite, but does so with a larger and irregular basal plane spacing. The oxidation of graphite results in the formation of epoxide groups, as well as C–OH and COOH groups. It is the presence of some of these moieties that allows GO to be dispersed in water, allowing for its use in waterborne formulations. Although GO does not possess the electrical properties of single-sheet graphene, it can be swelled in water, which allows for intercalation of hydrophilic polymer between GO sheets, resulting in a composite that can be coated to produce a continuous film. After coating it may be possible to chemically convert GO to a reduced graphite oxide (r-GO) with improved electrical conductivity. X-ray diffraction (XRD) is ideally suited to evaluate GO–polymer composite samples for evidence of intercalation or exfoliation of GO. Examples of GO–polymer analysis by XRD are presented, along with results that demonstrate the effect of relative humidity (RH) on neat GO. Knowing the ambient RH during XRD data collection was found to be important to correctly assess the extent of polymer intercalation within the GO lattice.

Numerical Analysis of the Diffusive Field around a Growing Protein Crystal in Microgravity

t is believed that a microgravity environment may maintain ideal depletion zones of protein (PDZ) and impurity (IDZ) around growing crystals and may contribute to growing high-quality crystals. This can lead to an X-ray diffraction data collection of higher resolution with lower mosaicity, because of the better internal order and fewer defects in the crystals when compared to ground-grown crystals. The extent of these depletion zones are dependent on a competition between the diffusion of the molecules in the solution (indexed by the diffusion coefficient, D) and the adsorption of those into the growing crystal (indexed by the kinetic constant, β). If we use the D/β value as an index of the extent of PDZ and IDZ, a lower D/β value is ideal for maintaining PDZ and IDZ. Using experimental results, we could easily obtain the D/β value. When we combined the D/β value with the quality of protein crystals obtained in microgravity experiments provided by Japanese Space Agency (JAXA), we found that the effects of microgravity contributed to obtaining superior crystals especially if the D/β value was less than 3 mm. The numerical analysis of the PDZ and IDZ shows that the radius of the crystal (R) is also related to the PDZ and the IDZ. If the Rβ/D value is large, both the PDZ and the IDZ provide a filtration effect, but if the Rβ/D value is small, only the IDZ does.

An in situ approach to study trace element partitioning in the laser heated diamond anvil cell

Data on partitioning behavior of elements between different phases at in situ conditions are crucial for the understanding of element mobility especially for geochemical studies. Here, we present results of in situ partitioning of trace elements (Zr, Pd, and Ru) between silicate and iron melts, up to 50 GPa and 4200 K, using a modified laser heated diamond anvil cell (DAC). This new experimental set up allows simultaneous collection of x-ray fluorescence (XRF) and x-ray diffraction (XRD) data as a function of time using the high pressure beamline ID27 (ESRF, France). The technique enables the simultaneous detection of sample melting based to the appearance of diffuse scattering in the XRD pattern, characteristic of the structure factor of liquids, and measurements of elemental partitioning of the sample using XRF, before, during and after laser heating in the DAC. We were able to detect elements concentrations as low as a few ppm level (2-5 ppm) on standard solutions. In situ measurements are complimented by mapping of the chemical partitions of the trace elements after laser heating on the quenched samples to constrain the partitioning data. Our first results indicate a strong partitioning of Pd and Ru into the metallic phase, while Zr remains clearly incompatible with iron. This novel approach extends the pressure and temperature range of partitioning experiments derived from quenched samples from the large volume presses and could bring new insight to the early history of Earth.

X-ray diffraction mapping on a curved surface

An efficient method for X-ray diffraction data collection mapping on a given curved surface has been developed. The method uses a laser–video autozalignment system to collect a map of heights on a fine mesh grid. It also reconstructs the surface geometry and determines surface normals on a three-dimensional surface fit. An algorithm was used to calculate the required rotation and tilt angles to coincide the sample normal with the diffraction center before each exposure. A set of diffraction frames collected using this technique was analyzed in order to superimpose a phase transformation map onto a typical zirconia ceramic sample. With this method, mapping of phase and stresses on a complex surface has been demonstrated. This approach is broadly applicable to many important areas of study.

Anomalous octahedral distortions in LaFe1−xNixO3

Perovskite-type LaFe1−xNixO3 in the composition range with orthorhombic crystal structure (x = 0.0, 0.1, 0.3 and 0.5) was prepared by the standard solid-state reaction method. The phase formation was checked by synchrotron x-ray diffraction. The structure was refined from the collected x-ray diffraction data using the Reitveld refinement method. The synthesized compositions crystallized mainly in the Pbnm symmetry. The available data in the literature on the structural properties and crystal phase evolution of the solid solution series LaFe1−xNixO3 were also analysed critically. We report an anomalous variation in the octahedral deformations in these compositions. In order to explore the composition-dependent microscopic changes driving these structural distortions, we used the refined structural parameters (cell dimensions and atomic coordinates) from our own results and the literature to calculate and see the effects of composition on several structural distortion parameters in an orthorhombic perovskite. Variation of these parameters strongly suggested that incorporation of Ni in LaFeO3 caused the octahedral deformation to increase. However, octahedral tilting decreased with the addition of Ni in LaFeO3. The obtained change in the orthorhombic distortions was caused by the possible presence of Jahn–Teller Fe4+ cations.

Use of chromophoric ligands to visually screen co-crystals of putative protein-nucleic acid complexes.

Distinguishing between crystals of protein-nucleic acid complexes and those containing protein alone is a common problem in structural studies of protein-nucleic acid interactions. Currently, there are several methods available for detecting nucleic acid in crystals, including gel electrophoresis, SYBR Gold fluorescence dye staining, and methyl violet staining. However, they require either that the crystals be sacrificed or access to a fluorescence microscope. In this protocol, we describe an approach that allows direct visualization of either the presence or absence of oligonucleotides in crystals grown from solutions containing both protein and nucleic acid--labeling with the Cy5 dye. In addition to offering the advantage of being able to distinguish between crystals of complex and protein alone with the naked eye or a light microscope, crystals of covalently Cy5-labeled DNA can be directly used for X-ray diffraction data collection.

Arabidopsis Chloroplastic Glutaredoxin C5 as a Model to Explore Molecular Determinants for Iron-Sulfur Cluster Binding into Glutaredoxins

Unlike thioredoxins, glutaredoxins are involved in iron-sulfur cluster assembly and in reduction of specific disulfides (i.e. protein-glutathione adducts), and thus they are also important redox regulators of chloroplast metabolism. Using GFP fusion, AtGrxC5 isoform, present exclusively in Brassicaceae, was shown to be localized in chloroplasts. A comparison of the biochemical, structural, and spectroscopic properties of Arabidopsis GrxC5 (WCSYC active site) with poplar GrxS12 (WCSYS active site), a chloroplastic paralog, indicated that, contrary to the solely apomonomeric GrxS12 isoform, AtGrxC5 exists as two forms when expressed in Escherichia coli. The monomeric apoprotein possesses deglutathionylation activity mediating the recycling of plastidial methionine sulfoxide reductase B1 and peroxiredoxin IIE, whereas the dimeric holoprotein incorporates a [2Fe-2S] cluster. Site-directed mutagenesis experiments and resolution of the x-ray crystal structure of AtGrxC5 in its holoform revealed that, although not involved in its ligation, the presence of the second active site cysteine (Cys(32)) is required for cluster formation. In addition, thiol titrations, fluorescence measurements, and mass spectrometry analyses showed that, despite the presence of a dithiol active site, AtGrxC5 does not form any inter- or intramolecular disulfide bond and that its activity exclusively relies on a monothiol mechanism.

Simultaneous X-ray diffraction from multiple single crystals of macromolecules

The potential in macromolecular crystallography for using multiple crystals to collect X-ray diffraction data simultaneously from assemblies of up to seven crystals is explored. The basic features of the algorithms used to extract data and their practical implementation are described. The procedure could be useful both in relation to diffraction data obtained from intergrown crystals and to alleviate the problem of rapid diffraction decay arising from the effects of radiation damage.

X-CHIP: an integrated platform for high-throughput protein crystallization and on-the-chip X-ray diffraction data collection.

The X-CHIP (X-ray Crystallization High-throughput Integrated Platform) is a novel microchip that has been developed to combine multiple steps of the crystallographic pipeline from crystallization to diffraction data collection on a single device to streamline the entire process. The system has been designed for crystallization condition screening, visual crystal inspection, initial X-ray screening and data collection in a high-throughput fashion. X-ray diffraction data acquisition can be performed directly on-the-chip at room temperature using an in situ approach. The capabilities of the chip eliminate the necessity for manual crystal handling and cryoprotection of crystal samples, while allowing data collection from multiple crystals in the same drop. This technology would be especially beneficial for projects with large volumes of data, such as protein-complex studies and fragment-based screening. The platform employs hydrophilic and hydrophobic concentric ring surfaces on a miniature plate transparent to visible light and X-rays to create a well defined and stable microbatch crystallization environment. The results of crystallization and data-collection experiments demonstrate that high-quality well diffracting crystals can be grown and high-resolution diffraction data sets can be collected using this technology. Furthermore, the quality of a single-wavelength anomalous dispersion data set collected with the X-CHIP at room temperature was sufficient to generate interpretable electron-density maps. This technology is highly resource-efficient owing to the use of nanolitre-scale drop volumes. It does not require any modification for most in-house and synchrotron beamline systems and offers a promising opportunity for full automation of the X-ray structure-determination process.