Mihoko Maruyama

Growth of Protein Crystals in Hydrogels Prevents Osmotic Shock

High-throughput protein X-ray crystallography offers a significant opportunity to facilitate drug discovery. The most reliable approach is to determine the three-dimensional structure of the protein-ligand complex by soaking the ligand in apo crystals. However, protein apo crystals produced by conventional crystallization in a solution are fatally damaged by osmotic shock during soaking. To overcome this difficulty, we present a novel technique for growing protein crystals in a high-concentration hydrogel that is completely gellified and exhibits high strength. This technique allowed us essentially to increase the mechanical stability of the crystals, preventing serious damage to the crystals caused by osmotic shock. Thus, this method may accelerate structure-based drug discoveries.

Enhancement of femtosecond laser-induced nucleation of protein in a gel solution

We found that the use of a gel solution with agarose enhanced femtosecond laser-induced nucleation and produced hen egg white lysozyme crystals at three to five times lower supersaturation than those by the femtosecond laser or agarose alone. The fast fluorescence imaging of the protein in the gel solution revealed that cavitation bubbles created high-concentration regions at the focal point, which could be the trigger for protein nucleation. The lower diffusions of protein molecules in agarose gel retained the high-concentration regions for a longer time, and facilitated the nucleation.

Fabrication of low-curvature 2 in. GaN wafers by Na-flux coalescence growth technique

Low-curvature and large-diameter GaN wafers are in high demand for the development of GaN-based electronic devices. Recently, we have proposed the coalescence growth of GaN by the Na-flux method and demonstrated the possibility of enlarging the diameter of high-quality GaN crystals. In the present study, 2 in. GaN wafers with a radius of curvature larger than 100 m were successfully produced by the Na-flux coalescence growth technique. FWHMs of the 002 and 102 GaN X-ray rocking curves were below 30.6 arcsec, and the dislocation density was less than the order of 102 cm−2 for the entire area of the wafer.

Promotion of Crystal Nucleation of Protein by Semi-Solid Agarose Gel

We investigate the nucleation rate of protein crystals in the presence of agarose gel at concentrations between 0.0 and 2.0% (w/v). It was found that agarose gel promoted the number of protein crystals up to a concentration of 0.4% (w/v), and gel at concentrations between 0.4 and 0.8% (w/v) decreased the nucleation rate. However, the number of crystals increased again in the presence of agarose by more than 1.0% (w/v), demonstrating high gel strength and a semi-solid gel. We propose that a crystallization method using semi-solid agarose is powerful for the screening of crystallization conditions.

Coalescence Growth of Dislocation-Free GaN Crystals by the Na-Flux Method

We have recently shown that dislocation-free GaN crystals could be grown on a GaN point seed by the Na-flux method. To enlarge the diameter of dislocation-free GaN crystals, we propose here the coalescence of GaN crystals grown from many isolated point seeds. In this study, we found that two GaN crystals grown from two point seeds arranged along the a-direction coalesced without generating dislocations at the coalescence boundary, and the c-axis misorientation between two crystals around the coalescence boundary gradually diminished as the growth proceeded. These results indicate that coalescence growth may become a key technique for fabricating large-diameter dislocation-free GaN crystals.

Growth of Prismatic GaN Single Crystals with High Transparency on Small GaN Seed Crystals by Ca–Li-Added Na Flux Method

The addition of Ca–Li to Na flux was attempted in order to control the growth habit and further improve transparency of GaN crystals. As a result, the growth habit changed to prism shape by the addition of Ca. Furthermore, we succeeded in growing prismatic GaN crystals with high transparency by adding appropriate amounts of Ca and Li to the flux. The optical absorption coefficient at 450 nm wavelength obtained from the crystal was 1.07 cm-1. This result suggests that the addition of Ca–Li to Na flux is a promising method of growing transparent GaN single crystals.

Approach for growth of high-quality and large protein crystals

Three crystallization methods, including crystallization in the presence of a semi-solid agarose gel, top-seeded solution growth (TSSG) and a large-scale hanging-drop method, have previously been presented. In this study, crystallization has been further evaluated in the presence of a semi-solid agarose gel by crystallizing additional proteins. A novel crystallization method combining TSSG and the large-scale hanging-drop method has also been developed.

Selective crystallization of metastable phase of acetaminophen by ultrasonic irradiation

A new method for selective crystallization of the metastable phase (form II) of acetaminophen is described. To obtain form II, we prepared a highly supersaturated solution (σI = 3.7) and then applied ultrasonic irradiation at different frequencies. Without ultrasonic irradiation, spontaneous crystallization did not occur within one month in the highly supersaturated condition (σI = 3.7). When ultrasonic irradiation at 28 kHz was applied, form II preferentially crystallized. Therefore, we conclude that ultrasonic irradiation can be an effective technique for selectively crystallizing the metastable phase.

Growth of bulk GaN crystals by the Na-flux point seed technique

In this paper, progress in the Na-flux point seed technique (SPST) will be reviewed. Bulk GaN crystals with a diameter of 2.1 cm, a height of 1.2 cm, and large dislocation-free areas were successfully produced by SPST. Panchromatic cathodoluminescence images of a wafer sliced parallel to the c-face from the crystal showed the lack of dark spots due to dislocations over a large area of the wafer. Structural properties were evaluated using synchrotron X-ray diffraction analysis at SPring-8. The full width at half maximum of the 006 rocking curve was found to be 2.1 arcsec, close to the calculated value of 2.0 arcsec for a perfect GaN crystal, indicating that crystals grown by SPST have an almost perfect structure. In addition, we have extended the use of SPST to the coalescence growth of GaN crystals to increase the wafer diameter and obtained a 2 in. GaN wafer with a low dislocation density and a low curvature by this technique.

Effects of Solution Stirring on the Growth of Bulk GaN Single Crystals by Na Flux Method

Recently, we succeeded in fabricating centimeter-sized bulk gallium nitride (GaN) crystals with large dislocation-free areas on a GaN point seed. However, problems of polycrystal formation, skeletal growth, and low growth rate still remained. In this study, to suppress skeletal growth, polycrystals formation and increase the growth rate, we introduced two types of solution-stirring techniques – rotating stirring and swinging stirring – in the growth on point seeds by the Na flux method. We found that increasing the reversal frequency of the rotating stirring and increasing the rate of the swinging stirring increased the growth rate and suppressed the formation of polycrystals and skeletal growth. Moreover, the maximum c-direction growth rate of 46 µm/h was achieved without the formation of polycrystals and skeletal growth. We conclude that solution stirring may be an effective technique for fabricating high-quality large bulk GaN crystals.