FNAS/advanced protein crystal growth

A scintillation method is presented for determination of the temperature dependence of the solubility, S(T), of proteins in 50-100 micro-l volumes of solution. S(T) data for lysozyme and horse serum albumin were obtained for various combinations of pH and precipitant concentrations. The resulting kinetics and equilibrium information was used for dynamic control, that is the separation of nucleation and growth stages in protein crystallization. Individual lysozyme and horse serum albumin crystals were grown in 15-20 micro-l solution volumes contained in x-ray capillaries

Mass spectroscopic facility

A mass spectrometer system is described. The investigations to be conducted with the completed facility will be concerned with the high-temperature out-gassing behavior of quartz ampoules and the vapor composition in crystal growth experiments. These will give some insight on the prevailing transport mechanisms in these crystal growth experiments and supply a realistic basis for the numerical modeling of heat and mass transfer in these systems

Novel protein crystal growth technology: Proof of concept

A technology for crystal growth, which overcomes certain shortcomings of other techniques, is developed and its applicability to proteins is examined. There were several unknowns to be determined: the design of the apparatus for suspension of crystals of varying (growing) diameter, control of the temperature and supersaturation, the methods for seeding and/or controlling nucleation, the effect on protein solutions of the temperature oscillations arising from the circulation, and the effect of the fluid shear on the suspended crystals. Extensive effort was put forth to grow lysozyme crystals. Under conditions favorable to the growth of tetragonal lysozyme, spontaneous nucleation could be produced but the number of nuclei could not be controlled. Seed transfer techniques were developed and implemented. When conditions for the orthorhombic form were tried, a single crystal 1.5 x 0.5 x 0.2 mm was grown (after in situ nucleation) and successfully extracted. A mathematical model was developed to predict the flow velocity as a function of the geometry and the operating temperatures. The model can also be used to scaleup the apparatus for growing larger crystals of other materials such as water soluble non-linear optical materials. This crystal suspension technology also shows promise for high quality solution growth of optical materials such as TGS and KDP

Fluid dynamics and low gravity effects of chemical vapor deposition

Based on the comparison between experimental data and numerical results for the growth of GaAs from TMGa, it was shown that 3D simulations are necessary to simulate rectangular CVD reactors even when operated under subcritical (Ra) conditions. The important points found are summarized in the three attached reprints. The experimental studies of mixed convection in horizontal channels have shown three regimes of high Ra (22,220) number flows. At Re = 18.5, the rolls develop very quickly, significantly modulating the axial velocity even before it reaches the beginning of the hot plate. A few centimeters downstream, the velocities become asymmetric about the vertical centerplane and at x = 12 cm, become unsteady. These asymmetries were predicted theoretically, but experimental evidence has not been published prior to this work. At Re = 36, the axial velocity is only slightly modified at x = 0. Although the flow remains steady and symmetric about the vertical centerplane, there is a small spatial oscillation in the velocities over the length of the channel. The period of this oscillation was around 5 cm. At Re = 54, the longitudinal rolls developed smoothly over a length of 30 cm, with no asymmetries, unsteadiness, or spatial oscillations. Comparison of numerical simulations of these flows to experiments has revealed the importance and difficulty of setting proper thermal boundary conditions on the sidewalls. Calculated flows and experimentally measured flows showed very similar profiles, but at different axial locations, with the rolls developing more rapidly in the experiments. This is directly attributable to partially conducting sidewalls of the apparatus being hotter in the entrance section than the adiabatic walls of the simulations. A thorough comparison of the experimental data and numerical results for a variety of sidewall boundary conditions is in preparation

Process modelling for materials preparation experiments

The main goals of the research under this grant consist of the development of mathematical tools and measurement of transport properties necessary for high fidelity modeling of crystal growth from the melt and solution, in particular, for the Bridgman-Stockbarger growth of mercury cadmium telluride (MCT) and the solution growth of triglycine sulphate (TGS). Of the tasks described in detail in the original proposal, two remain to be worked on: (1) development of a spectral code for moving boundary problems; and (2) diffusivity measurements on concentrated and supersaturated TGS solutions. Progress made during this seventh half-year period is reported

Nucleation and Convection Effects in Protein Crystal Growth

Work during the second year under this grant (NAG8-1161) resulted in several major achievements. We have characterized protein impurities as well as microheterogeneities in the proteins hen egg white lysozyme and horse spleen apoferritin, and demonstrated the effects of these impurities on nucleation and crystallization. In particular, the purification of apoferritin resulted in crystals with an X-ray diffraction resolution of better than 1.8 A, i.e. a 1 A improvement over earlier work on the cubic form. Furthermore, we have shown, in association with studies of liquid-liquid phase separation, that depending on the growth conditions, lysozyme can produce all growth morphologies that have been observed with other proteins. Finally, in connection with our experimental and simulation work on growth step bunching, we have developed a system-dependent criterion for advantages and disadvantages of crystallization from solution under reduced gravity. In the following, these efforts are described in some detail

Morphological stability and kinetics in crystal growth from vapors

The following topics are discussed: (1) microscopy image storage and processing system; (2) growth kinetics and morphology study with carbon tetrabromide; (3) photothermal deflection vapor growth setup; (4) bridgman growth of iodine single crystals; (5) vapor concentration distribution measurement during growth; and (6) Monte Carlo modeling of anisotropic growth kinetics and morphology. A collection of presentations and publications of these results are presented

Convective flow effects on protein crystal growth

During the fifth semi-annual period under this grant we have pursued the following activities: (1) Characterization of the purity and further purification of lysozyme solutions, these efforts are summarized in Section 2; (2) Crystal growth morphology and kinetics studies with tetragonal lysozyme, our observation on the dependence of lysozyme growth kinetics on step sources and impurities has been summarized in a manuscript which was accepted for publication in the Journal of Crystal Growth; (3) Numerical modelling of the interaction between bulk transport and interface kinetics, for a detailed summary of this work see the manuscript which was accepted for publication in the Journal of Crystal Growth; and (4) Light scattering studies, this work has been summarized in a manuscript that has been submitted for publication to the Journal of Chemical Physics

Growth of zinc selenide crystals by physical vapor transport in microgravity

The growth of single crystals of zinc selenide was carried out by both closed ampoule physical vapor transport and effusive ampoule physical vapor transport (EAPVT). The latter technique was shown to be a much more efficient method for the seeded growth of zinc selenide, resulting in higher transport rates. Furthermore, EAPVT work on CdTe has shown that growth onto /n11/ seeds is advantageous for obtaining reduced twinning and defect densities in II-VI sphalerite materials

Research support for cadmium telluride crystal growth

The growth of single crystals of zinc selenide was carried out by both closed ampoule physical vapor transport and effusive ampoule physical vapor transport (EAPVT). The latter technique was shown to be a much more efficient method for the seeded growth of zinc selenide, resulting in higher transport rates. Furthermore, EAPVT work on CdTe has shown that growth onto (n 11) seeds is advantageous for obtaining reduced twinning and defect densities in II-VI sphalerite materials