In situ X-ray analysis of protein crystals in low-birefringent and X-ray transmissive plastic microchannels

Plastic microchannel crystallization template designs made from inexpensive cyclic olefin copolymers have been shown to be low‐birefringent, X‐ray transmissive and compatible with microfluidic fabrication in restricted geometry. The model proteins thaumatin, lysozyme and bacteriorhodopsin demonstrat...

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Bibliographic Details
Published in:Acta crystallographica. Section D, Biological crystallography. Biological crystallography., 2008-02, Vol.64 (2), p.189-197
Main Authors: Ng, Joseph D., Clark, Peter J., Stevens, Raymond C., Kuhn, Peter
Format: Article
Language:English
Subjects:
Bacteriorhodopsins - chemistry
Crystallization
Crystallography, X-Ray - instrumentation
Crystallography, X-Ray - methods
in situ X-ray analysis
microchannels
Muramidase - chemistry
Plant Proteins - chemistry
Proteins - chemistry
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Summary:Plastic microchannel crystallization template designs made from inexpensive cyclic olefin copolymers have been shown to be low‐birefringent, X‐ray transmissive and compatible with microfluidic fabrication in restricted geometry. The model proteins thaumatin, lysozyme and bacteriorhodopsin demonstrated the feasibility of conducting counter‐diffusion equilibration within the new plastic configuration. Crystals of each of these proteins were directly evaluated in situ using synchrotron radiation and their diffraction quality was evaluated without invasive manipulation or cryofreezing. Protein crystals able to produce complete X‐ray data sets were used to calculate electron‐density maps for structure determination. Fluidic crystallization in the plastic platform was also coupled with a commercialized automated imager and an in situ X‐ray scanner that allowed optical and X‐ray inspection of crystallization hits. The results demonstrate the feasibility of rapid nanovolume counter‐diffusion crystallization experiments without the need for additional instrumentation.
ISSN:1399-0047
0907-4449
1399-0047
DOI:10.1107/S0907444907060064