Microgravity Crystallization Research Articles

Preparation of Experiments on Growing Zinc–Cadmium Telluride Crystals in Microgravity

Cd1-xZnxTe crystals are necessary for the production of ionizing radiation detectors widely used in science, technology, medicine and other fields. Internal stresses during crystallization lead to generation of dislocations and low-angle boundaries. Typical problem of melt crystal growth of Cd-Zn-Te compounds are tellurium inclusions, which deteriorate detector performance. Microgravity conditions provide unique opportunities for growing high-quality crystals due to the absence of convection, more equilibrium conditions of melt mixing, and a decrease in internal stresses. Since the properties of such crystals strongly depend on the production conditions, seeds and a feed ingot with specified compositions and structure are required. Ampoules with two compositions of materials have been prepared for the space experiment. Crystals of different compositions Cd0.96Zn0.04Te and Cd0.9Zn0.1Te were produced for two charges. They consist of an oriented seed, solvent, and feeding ingot, which are single-phased, single crystalline, have certain crystallographic orientation, meet demands for growth of Cd–Zn–Te crystals in microgravity. Ampoules containing these materials were sent to International Space Station for crystal growth on equipment already assembled at “Nauka” station.

Microgravity crystallization and neutron diffraction of perdeuterated tryptophan synthase

Microgravity crystallization and neutron diffraction of perdeuterated tryptophan synthase

Microgravity crystallization of perdeuterated tryptophan synthase for neutron diffraction

Biologically active vitamin B6-derivative pyridoxal 5′-phosphate (PLP) is an essential cofactor in amino acid metabolic pathways. PLP-dependent enzymes catalyze a multitude of chemical reactions but, how reaction diversity of PLP-dependent enzymes is achieved is still not well understood. Such comprehension requires atomic-level structural studies of PLP-dependent enzymes. Neutron diffraction affords the ability to directly observe hydrogen positions and therefore assign protonation states to the PLP cofactor and key active site residues. The low fluxes of neutron beamlines require large crystals (≥0.5 mm3). Tryptophan synthase (TS), a Fold Type II PLP-dependent enzyme, crystallizes in unit gravity with inclusions and high mosaicity, resulting in poor diffraction. Microgravity offers the opportunity to grow large, well-ordered crystals by reducing gravity-driven convection currents that impede crystal growth. We developed the Toledo Crystallization Box (TCB), a membrane-barrier capillary-dialysis device, to grow neutron diffraction-quality crystals of perdeuterated TS in microgravity. Here, we present the design of the TCB and its implementation on Center for Advancement of Science in Space (CASIS) supported International Space Station (ISS) Missions Protein Crystal Growth (PCG)-8 and PCG-15. The TCB demonstrated the ability to improve X-ray diffraction and mosaicity on PCG-8. In comparison to ground control crystals of the same size, microgravity-grown crystals from PCG-15 produced higher quality neutron diffraction data. Neutron diffraction data to a resolution of 2.1 Å has been collected using microgravity-grown perdeuterated TS crystals from PCG-15.

Tracing transport of protein aggregates in microgravity versus unit gravity crystallization

Microgravity conditions have been used to improve protein crystallization from the early 1980s using advanced crystallization apparatuses and methods. Early microgravity crystallization experiments confirmed that minimal convection and a sedimentation-free environment is beneficial for growth of crystals with higher internal order and in some cases, larger volume. It was however realized that crystal growth in microgravity requires additional time due to slower growth rates. The progress in space research via the International Space Station (ISS) provides a laboratory-like environment to perform convection-free crystallization experiments for an extended time. To obtain detailed insights in macromolecular transport phenomena under microgravity and the assumed reduction of unfavorable impurity incorporation in growing crystals, microgravity and unit gravity control experiments for three different proteins were designed. To determine the quantity of impurity incorporated into crystals, fluorescence-tagged aggregates of the proteins (acting as impurities) were prepared. The recorded fluorescence intensities of the respective crystals reveal reduction in the incorporation of aggregates under microgravity for different aggregate quantities. The experiments and data obtained, provide insights about macromolecular transport in relation to molecular weight of the target proteins, as well as information about associated diffusion behavior and crystal lattice formation. Results suggest one explanation why microgravity-grown protein crystals often exhibit higher quality. Furthermore, results from these experiments can be used to predict which proteins may benefit more from microgravity crystallization.

Erratum to: Crystallization in Microgravity and the Atomic-Resolution Structure of Uridine Phosphorylase from Vibrio cholerae

Erratum to: Crystallization in Microgravity and the Atomic-Resolution Structure of Uridine Phosphorylase from Vibrio cholerae

Microgravity crystallization for improving uniformity and homogeneity of crystals for time-resolved diffusive mixing XFEL experiments

Microgravity crystallization for improving uniformity and homogeneity of crystals for time-resolved diffusive mixing XFEL experiments

Technology of high quality protein crystals’ obtaining aboard the ISS at execution of joint Japanese–Russian experiments

A series of space experiments aboard the International Space Station (ISS) associated with high-quality Protein Crystal Growth (PCG) in microgravity conditions can be considered as a unique and one of the best examples of fruitful collaboration between Japanese and Russian scientists and engineers in space, which includes also other ISS International Partners. X-ray diffraction is still the most powerful tool to determine the protein three dimensional structure necessary for Structure based drug design (SBDD). The major purpose of the experiment is to grow high quality protein crystals in microgravity for X-ray diffraction on Earth. Within one and a half decade, Japan and Russia have established an efficient process over PCG in space to support latest developments over drug design and structural biology. One of the keys for success of the experiment lies in how precisely pre-launch preparations are made. Japanese party provides flight equipment for crystallization and ensures the required environment to support the experiment aboard of the ISS’s Kibo module, and also mainly takes part of the experiment ground support such as protein sample characterization, purification, crystallization screening, and solution optimization for microgravity experiment. Russian party is responsible for integration of the flight items equipped with proteins and precipitants on board Russian transportation space vehicles (Soyuz or Progress), for delivery them at the ISS, transfer to Kibo module, and returning the experiments’ results back on Earth aboard Soyuz manned capsule. Due to close cooperation of the parties and solid organizational structure, samples can be launched at the ISS every half a year if the ground preparation goes smoothly. The samples are crystallized using counter diffusion method at 20 degree C for 1–2.5 months. After samples return, the crystals are carefully taken out from the capillary, and frozen for X-ray diffraction at SPring8 facility in Japan. Extensive support of researchers from both countries is also a part of this process. The paper analyses details of the PCG experiment scheme, unique and reliable technology of its execution, and contains examples of the application. Key words: International Space Station, Protein crystals, Microgravity, International collaboration.

Microgravity protein crystallization for drug development: a bold example of public sector entrepreneurship

A basic mission of NASA is to use the United States’ segment of the International Space Station (ISS), designated a national laboratory, to facilitate the growth of a commercial marketplace in low Earth orbit for scientific research, technology development, observation and communications. Protein crystallization research has long been promoted as a promising commercial application of the ISS for drug development. In this paper we examine the case for microgravity protein crystallization under different private and public investment scenarios. The analysis suggests that sustaining investment is unlikely to come from individual companies alone. Public and private investment must be combined and managed to overcome a number of challenges including the need to integrate microgravity crystallization into the complex system of technologies involved in structure-based drug design. Multiple risks related to transportation costs/frequency, risk for cargo and research crew, and uncertainty about the longevity of the ISS complicate the calculus.

Effects of microgravity crystallization on a ligand-induced RNA crystal phase transition

Effects of microgravity crystallization on a ligand-induced RNA crystal phase transition

Implementation of Temperature-Controlled Method of Protein Crystallization in Microgravity

An experimental scientific equipment for implementing temperature-controlled protein crystallization in capillaries under microgravity has been developed, fabricated, and tested. This crystallization method, providing on-line separate control of crystal growth both in the stage of nucleation of crystals and during their further growth, requires small amounts of protein solution. The equipment has been tested on board of Foton-M4 spacecraft (growth of lysozyme protein crystals of high structural quality in microgravity) using a cyclogram developed in ground-based experiments. The results obtained have demonstrated efficiency and importance of the developed equipment and method for growing biomacromolecular crystals of high-structural quality.

Experimental and Analytical Techniques for Studying ZBLAN Crystallization in Microgravity

One of the promising new areas of materials research is in the field of microgravity. Microgravity experimentation enables new materials to be developed and traditional materials to be improved, which cannot be completed under terrestrial conditions. Recent experiments on ZBLAN (ZrF4–BaF2–LaF3–AlF3 –NaF) glass have shown that, when heated, there is a crystallization dependency on gravity. This crystallization dependency limits the optical transmissibility of this material, due to crystallites forming during the fiber drawing process. ZBLAN glass has the theoretical potential for optical transmission in the range of 0.3 –7 μm, which would facilitate much needed mid-infrared (IR) fiber technology. Past researchers have completed ZBLAN crystallizationmicrogravity experiments, with limited details on the experimental technique and analysis methods. This study demonstrates an alternative experimental technique for ZBLAN microgravity testing and postprocessing techniques that reveal crystallinity in the sample.

New scientific equipment for protein crystallization in microgravity, BELKA, and its approbation on the Bion-M No. 1 spacecraft

A space experiment on the crystallization of lisozyme and glucose isomerase proteins in UK-1 and UK-2 crystallizers on the scientific equipment BELKA on the Bion-M no. 1 spacecraft was performed in April–May 2013. A ground-based experiment was carried out simultaneously at the Institute of Crystallography of the Russian Academy of Sciences (IC RAS). Transparent crystals were obtained in both cases. The lisozyme crystals grown in microgravity are larger than their terrestrial analogs. An optical study of glucose isomerase crystals grown in space has shown that the coalescence of equally oriented crystallites leads to the formation of quasi-single-crystal blocks. An X-ray diffraction experiment on lisozyme crystals has revealed the resolutions for crystals obtained under terrestrial conditions and in space to be 1.74 and 1.58 A, respectively.

Growth of a Si0.50Ge0.50 crystal by the traveling liquidus-zone (TLZ) method in microgravity

An alloy semiconductor Si1−xGex (x~0.5) crystal was grown by the TLZ method in microgravity. Ge concentration was 48.5±1.5at% for the whole region of 10mm diameter and 17.2mm long crystal. Compositional uniformity was established but the average concentration was a little deviated from the expected 50at%. For further improving compositional uniformity and for obtaining Si0.5Ge0.5 crystals in microgravity, growth conditions were refined based on the measured axial compositional profile. In determining new growth conditions, difference in temperature gradient in a melt, difference in freezing interface curvature, and difference in melt back length of a seed between microgravity and terrestrial growth were taken into consideration.

Precise Measurements of Dendrite Growth of Ice Crystals in Microgravity

An ice crystal growth experiment was performed on board the International Space Station. The experiment was repeated 134 times with various undercooling conditions. Dendrite crystal growth velocity and tip radius were precisely measured by using a newly developed software. The results are compared with those obtained previously on the ground as well as with those reported by Glicksman et al. for Succinonitrile (SCN). The plot of the dimensionless velocity V as a function of the dimensionless undercooling Δ under microgravity revealed values that were consistently lower than those obtained under 1-G which indicates that thermal convection was suppressed. The plot of the dimensionless radius R0 as a function of Δ showed a less scattered value. The stability factor σ* became close to that of SCN when it was calculated using the geometrical mean radius.

Crystallization in gels and microgravity: a comparative study

Crystallization in gels and microgravity: a comparative study

Stability of magnetically suppressed solutal convection in crystal growth from solutions

Convective motions during different crystal growth processes are known to interfere with the quality of the growth process and are hypothesized to be detrimental to the quality of the grown crystal. In crystal growth from solutions for example, without fluid interfaces and free surfaces, gravity driven phenomena such as thermal and solutal buoyancy effects and sedimentation are the primary causes of fluid flows in the system. While crystallization in microgravity can approach diffusion limited growth conditions (no convection), terrestrially strong magnetic fields can be used to control fluid flow and sedimentation effects. In this work, a theory is presented on the stability of solutal convection of a magnetized fluid in the presence of a magnetic field. The requirements for stability are developed and compared to experiments performed within the bore of a superconducting magnet. The theoretical predictions are in good agreement with the experiments and show solutal convection can be stabilized if the surrounding fluid has larger magnetic susceptibility and the magnetic field has a specific structure. Discussion on the application of the technique to protein crystallization is also provided.

Macromolecular Crystallization in Microgravity Generated by a Superconducting Magnet

About 30% of the protein crystals grown in space yield better X-ray diffraction data than the best crystals grown on the earth. The microgravity environments provided by the application of an upward magnetic force constitute excellent candidates for simulating the microgravity conditions in space. Here, we describe a method to control effective gravity and formation of protein crystals in various levels of effective gravity. Since 2002, the stable and long-time durable microgravity generated by a convenient type of superconducting magnet has been available for protein crystal growth. For the first time, protein crystals, orthorhombic lysozyme, were grown at microgravity on the earth, and it was proved that this microgravity improved the crystal quality effectively and reproducibly. The present method always accompanies a strong magnetic field, and the magnetic field itself seems to improve crystal quality. Microgravity is not always effective for improving crystal quality. When we applied this microgravity to the formation of cubic porcine insulin and tetragonal lysozyme crystals, we observed no dependence of effective gravity on crystal quality. Thus, this kind of test will be useful for selecting promising proteins prior to the space experiments. Finally, the microgravity generated by the magnet is compared with that in space, considering the cost, the quality of microgravity, experimental convenience, etc., and the future use of this microgravity for macromolecular crystal growth is discussed.

Upside‐Down Protein Crystallization

The benefits of protein crystal growth in microgravity are well documented. The crystallization vessels currently employed for microgravity crystallization are far from optimal with regards to cost, sample volume, size, and ease of use. The use of microbatch experiments is a favorable alternative in each respect: 96 experiments of 0.5-2 microL volumes can be performed in a single microtiter tray measuring 5 x 8 cm and costing 1 pound sterling each. To date, the use of microbatch has not been pursued on account of concerns of oil leakage. To address this issue, a novel approach to microbatch crystallization experiments is described, where the microbatch plates are inverted throughout the duration of the experiment. The findings intimate the application of the microbatch method to space flight and the potential to drastically increase the output of microgravity crystallization research .

Excellent compositional homogeneity in In0.3Ga0.7As crystals grown by the traveling liquiduszone (TLZ) method

The influence of convection in a melt on the compositional homogeneity of the TLZ-grown In0.3Ga0.7As crystals has been investigated by growing crystals with various dimensions on the ground. Excellent compositional homogeneity such as 0.3 plus or minus 0.01 in InAs mole fraction for a distance of 25 mm was obtained when the melt diameter was limited to 2 mm and convective flow in the melt was suppressed. On the other hand, when the crystal diameter was increased to 10 mm, both axial and radial compositional homogeneity was deteriorated due to convection in the melt. Comparing with the numerical simulation, convective flow velocity less than 1.4 mm/h may be sufficient for growing homogeneous crystals and it is not so difficult to suppress convective flow velocity below 1.4 mm/h for 10 mm diameter crystals in microgravity. Therefore, larger homogeneous In0.3Ga0.7As crystals are expected to be grown by the TLZ method on board the International Space Station.

Crystallization Trials of Nitroreductase NfsB on the Ground for the Application to Crystallization in Microgravity

Crystallization Trials of Nitroreductase NfsB on the Ground for the Application to Crystallization in Microgravity