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Manipulation and growth of birefringent protein crystals in optical tweezers

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Abstract

We report on the trapping, rotation, and in-situ growth of birefringent tetragonal lysozyme crystals in optical tweezers operating at a wavelength of 1070 nm. Variation of the pH and lysozyme concentration of the solution during growth was used to alter the length to width ratio of the crystals, and hence their orientation in the tweezers. Crystals with the optical axis skewed or perpendicular to the trapping-beam axis could be rotated by changing the orientation of linearly polarized light. We observed spontaneous spinning of some asymmetric crystals in the presence of linearly polarized light, due to radiation pressure effects. Addition of protein to the solution in the tweezers permitted real-time observation of crystal growth.

©2004 Optical Society of America

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Figures (6)

Fig. 1.
Fig. 1. Morphology of tetragonal lysozyme, showing crystallographic axes and faces. The optical axis coincides with the c-axis. The aspect ratio is defined as l:w.
Fig. 2.
Fig. 2. The optical tweezers setup used for trapping and imaging lysozyme crystals.
Fig. 3.
Fig. 3. Images of lysozyme crystals (pH 7.0, 1:20 protein:salt solution) taken in the tweezers setup under a) crossed polarizers and b) linearly polarized light. The polarizer axes are aligned with the image edges.
Fig. 4.
Fig. 4. Measured and calculated values of the momentum transfer vs. angle for several crystals (solid line is for Δn=1.66×10-3). Each symbol represents a distinct crystal.
Fig. 5.
Fig. 5. Movie of autorotation of a compound lysozyme crystal in a linearly polarized trapping beam. Rotation continued for ~10 minutes before slowing and stopping. (730 kB)
Fig. 6.
Fig. 6. Growth of lysozyme seed (from pH 7, 1:4 protein:salt solution) in the tweezers after addition of protein concentrate. Total elapsed time is 1 minute 12 seconds. (567 kB movie)
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