Dr. Muralidhar Krishnamurthy Profile

Dr. Muralidhar Krishnamurthy

at Indian Institute of Technology Kanpur

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Proceedings Article | 2 May 2008 Paper

Imaging and interferometric analysis of protein crystal growth

Ranjini Raghunandan, Anamika Sethia Gupta, K. Muralidhar

Proceedings Volume 6991, 69912H (2008) https://doi.org/10.1117/12.782119

KEYWORDS: Crystals, Proteins, Diffusion, Interferometry, Crystallography, Sodium, Mach-Zehnder interferometers, Molecules, CCD cameras, Liquid crystals

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Protein crystals are grown under controlled temperature, concentration and vapor pressure conditions, usually by vapor diffusion, liquid-liquid diffusion and dialysis techniques. The present study examines the effects of protein concentration, drop size and reservoir height on the crystal growth of Hen Egg White Lysozyme (HEWL). Crystals are grown by the hanging drop vapor diffusion method using Modular VDX^TM Plates. Due to the vapor pressure difference created between the protein drop and the reservoir, evaporation takes place till equilibrium is attained. Crystal formation takes place after a certain level of supersaturation is attained when the protein precipitates out in crystalline form. The observations revealed that the growth is faster for higher lysozyme concentration, smaller drop sizes and larger reservoir heights. The morphology of the crystals is viewed during the growth process using stereomicroscope. The number of crystals formed is the maximum for higher concentrations, drop sizes and reservoir heights. When the number of crystals formed is less, the size of the crystals is comparatively larger. The effect of evaporation of water vapor from the protein drop into the reservoir is studied using Mach-Zehnder interferometry. The recorded interferograms and shadowgraph images indicate the diffusion of condensed water into the reservoir. The radius of the drop is determined using the shadowgraph images of the growth process. The radius decreases with evaporation and the rate of decrease of radius is highest for higher protein concentrations, smaller drop sizes and larger reservoir heights.

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