Protein Crystallization Platform

The laboratory of Chemistry of Biological Processes is equipped with state of the art devices for the crystallization of biological macromolecules. The laboratory houses a Mosquito® Crystal (TTPLabtech) that allows the automated screening of crystallization conditions using low sample volumes and zero cross-contaminations. The robot allows the setup of sitting and hanging drop vapor diffusion trials and can perform microbatch under oil if desired. Our laboratory also owns a 5-head Dragonfly® (TTPLabtech) as a companion for the Mosquito®, which allows the setup of crystallizations screens for crystal optimization. To check the crystallization experiments, two stereomicroscopes are available including a LEICA M205C equipped with a camera IC80 HD. In addition, a home-made automated visualization robot adapted from Dr. Andrew Bohm’s design is available. Drop results analysis is done with our open-source software “AMi_Image_Analysis”.

The laboratory is interested in solving the structure of proteins with [Fe-S] clusters that are extremely sensitive to oxygen. To this end, we host a dedicated setup for protein crystallization in anaerobic conditions. It comprises two glove boxes including one containing an OryxNano (Douglas Instruments) that allows the screening of crystallization conditions with little sample volumes. The OryxNano can perform sitting-drop and microbatch experiments under oil. The hygrometry in the system is regulated between 45-50%. The crystallization drops are observed with a stereomicroscope LEICA M80 and pictures taken with a camera IC80 HD.

The laboratory is also well equipped for biophysical characterization of the samples with a circular dichroism spectrometer (ChirascanTM, Applied photophysics) useful for controlling the protein fold and measuring its thermal stability. A Dynamic light scattering instrument (Malvern Zetasizer ZSP) allows the estimation of the polydispersity of the sample and its stability, the calculation of the hydrodynamic radius as well as the measurement of the molecular weight and second virial coefficient (A2). A fluorescence spectrophotometer is also available.

Les robots Mosquito® et Dragonfly®

The Mosquito® and Dragonfly® liquid handlers

Le stéréomicroscope LEICA M205C

The LEICA stereomicroscope

The AMi automated microscope

The AMi automated microscope

Boîtes à gants - 1

The gloveboxes for crystallization in anaerobic conditions

Boîtes à gants - 2

The gloveboxes for crystallization in anaerobic conditions

Spectromètre de dichroisme circulaire

The Circular Dichroism spectrometer (ChirascanTM , Applied photophysics)

Appareil de diffusion dynamique de la lumière

Dynamic Light Scattering instrument (Malvern Zetasizer ZSP)