The high throughput crystallization platform offers vapour diffusion as well as crystallization under oil screening and optimization in sitting drops. Flexible methods have been developed to adapt each step to individual project requirements. At the crystallization level, we have especially focused on developing techniques to optimize crystal growth.
Crystallisation is performed using general screens from commercial sources as well as specific screens made from our own stock solutions.
Sample characterization is a prerequisite before crystallisation. Unless it has already been performed, samples will be analyzed 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 optimization.
The process is automated at different steps:
- Home made and optimization screens are prepared with a Tecan Evo Workstation,
- Sitting drops -from 100nl to 500nl- are performed by a Mosquito robot,
- Batch crystallization 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°, 20° and 26°C and inspected through a Leica stereo-microscope.
Cristals 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 Pilatus 300K pixel array detector.