Service / Technology Instance

About Macromolecular Crystallisation, Strasbourg, France

View All Crystallisation at Instruct

The high throughput crystallisation platform offers vapour diffusion as well as crystallisation under oil screening and optimisation in sitting drops. Flexible methods have been developed to adapt each step to individual project requirements. At the crystallisation level, we have especially focused on developing techniques to optimise crystal growth.

Instruments Available:

The high throughput crystallisation platform offers vapour diffusion as well as crystallisation under oil screening and optimisation in sitting drops. Flexible methods have been developed to adapt each step to individual project requirements. At the crystallisation level, we have especially focused on developing techniques to optimise crystal growth. Crystallisation is performed using general screens from commercial sources as well as specific screens made from our own stock solutions. Hanging drop plates and capillaries for free interface diffusion are also proposed for manual screening and optimisation. The process is automated at different steps; Home made and optimisation screens are prepared with a Tecan Evo Workstation, Sitting drops, from 100nl to 500nl, are performed by a Mosquito robot, Batch crystallisation in microfluidics chip are set up using a Plugmaker Microcapillary Protein Crystallization System™, Plates are stored and automatically imaged at 4°C and 20°C in Rock Imager systems (Formulatrix). Images are accessible to users and can be scored via a web-interface. Plates can also be stored at 4°C, 20°C and 26°C and inspected through a Leica stereo-microscope.

Ultra high-intensity microfocus rotating anode X-ray generator coupled with a diffractometer for collection of diffraction data from single crystals, and a BioSAXS device for collection of diffusion data from macromolecular solutions. The variable divergence on the optics, coupled with a pixel array detector, allows for the resolution of unit cell dimensions up to 600Å.

User Guide

Crystallisation is performed using general screens from commercial sources as well as specific screens made from stock solutions. Sample characterisation is a prerequisite for crystallisation. Unless it has already been performed, samples will be analysed by native and SDS-PAGE, as well as dynamic light scattering and thermal shift assay. The platform also offers analysis by analytical ultracentrifugation. Hanging drop plates and capillaries for free interface diffusion are also proposed for manual screening and optimisation.

The process is automated at different steps:

  • Home made and optimisation screens are prepared with a Tecan Evo Workstation,
  • Sitting drops -from 100nl to 500nl- are performed by a Mosquito robot,
  • Batch crystallisation in microfluidics chip are set up using a Plugmaker Microcapillary Protein Crystallization System™,
  • Plates are stored and automatically imaged at 4°C and 20°C in Rock Imager systems (Formulatrix). Images are accessible to users and can be scored via a web-interface. Plates can also be stored at 4°C, 20°C and 26°C and inspected through a Leica stereo-microscope.

Crystals can be tested with our in-house X-ray source Rigaku FR-X generator coupled with Osmic VariMax HF Arc)Sec Confocal optics and a Dectris Eiger 4M pixel array detector.

Instruct Centre

Instruct Centre FR1

IGBMC

1, rue Laurent Fries

Illkirch

Strasbourg

France

www.igbmc.fr

Macromolecular Crystallisation, Strasbourg, France

Contacts:

Marie-Christine Poterszman
Marie-Christine Poterszman
CNRS
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Pierre Poussin-Courmontagne
Pierre Poussin-Courmontagne
IGBMC-CERBM
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Alastair McEwen
Alastair McEwen
IGBMC-CERBM
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