In situ observation of protein crystallization under high pressure

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1. Designing and build-in of the high pressure in situ observation system :A diamond anvil cell was designed to fit the metallurgical phase-contrast microscope. The pressure measurements system was built up using the fluorescence shift of a spectrum from a ruby laser beam.2. A possible way of controlling the number of nuclei by applying pressure on HEW lysozyme crystals was found.It was found that the number of nuclei of HEW lysozyme diminishes when the nucleation took place under pressure. The higher the pressure the less number of nuclei appeared.3. The pressure dependence of the solubility of HEW lysozyme :The pressure was changed from 510MPa to 0.1 MPa at 25(〕SY.+-.〔)0.2ﾟC using the anvil. It was found that the single crystals of HEW lysozyme (tetragonal system) grew as the pressure was decreased. It became clear for the first time that the solubility of HEW lysozyme increased when the pressure in the vessel was increased.4. Habit change of HEW lysozyme by applying pressure :The ratio of growth rate vertical to the faces (101) and (110) increased monotonously as the pressure was increased. This resulted in the habit change of the crystal ; the bulk crystals c axis.