CRYSTALLISATION
Paris Sud PF ST1 Crystallization
The Crystallization platform at South Paris (labelled IBiSA) allows screening, analysis and automated optimization of the macromolecules crystallization conditions by vapour diffusion methods at 20°C (soon also at 4°C). Until construction and installation of the new Institute for Integrative Biology of the Cell (I2BC) in a near future, this platform is located in two sites: LEBS (CNRS, Gif-sur-Yvette) and IBBMC (University of South Paris, Orsay).
In particular, it offers self-service access to crystallisation and visualization robots and provides users with ready to use commercial kits of crystallisation and home-made standard stocks solutions for crystal optimization, and can realize crystallography projects in collaboration.
Plates can be stored in 4°C and 20°C and followed with stereo-microscopes (Nikon. Leica)
The Crystallization platform disposes of following devices:
- Tecan Genesis and Freedom Evo(TECAN) Three stations, one acquired with support of FRISBI. Equipped with eight needles are used to dispense mother liquor solutions in the crystallisation reservoir (70-150µl) for screening and perform crystallization drops of large volume (1µl to 2µl).
- Cartesian Microsys(PROTEIGEN).Two stations used to set up low volume crystallization drops in prefilled plates by the Tecan robot. Equipped with eight needles, they allow indeed dispensing volumes between 50 and 300 nl. Protein consumption is reduced (between 13µl and 37µl of protein for 96 drops of a single concentration).
- Mosquito®(TTP LabTech). This robot is used to dispense nanolitre volumes, making using disposable positive displacement pipettes to avoid cross contamination. The robot automates protein crystallography set-ups and additive screening. This provides an ideal setting for our systematic proteolytic cleavage during crystallization strategy (Collinetet al., 2011).
- RockImager 182(Formulatrix). Crystallization cabinet (at 20°C) equipped with a robotized visualization system that at regular intervals takes pictures of the crystallisation drops and stores them on computer for inspection. TwoRockImager 1000will be soon installed thanks to FRISBI support. One of them to follows crystallization trials at 4°C and the other at 20°C with UV Imaging Options, which allow the detection of tryptophan and to distinguish protein from salt crystals.