Overview

The High Pressure Freezing Laboratory for Macromolecular Crystallography (HPMX) is a service offered to ESRF users as a complement to diffraction experiments on the Structural Biology beamlines. The laboratory uses gases at high pressuresto either allow cryo-protectant free cooling of macromolecular samples or the introduction of gases in macromolecules. The laboratory is equipped with and provides different types of pressure cells to introduce gases (e.g. noble gases, oxygen, CO2 etc.) in crystals to answer specific scientific questions in structural biology.

As mentioned above, we also offer to prepare and freeze cryopotectant-free crystals of macromolecules or complexes thereof, under He or Ar pressure  at 200 to 2000 bar  (see figure below) prior to cryo-cooling and subsequent collection of diffraction data. This high pressure treatment is an alternative to the classical  preparation of samples using cryprotectant prior to cryo-cooling at home laboratories. The HPMX laboratory is located within the MX village and in close proximity of ID30A-3 in the room 30-0-08.

 

Phase diagram of water

 

 

Equipment

 

Equipment which is available for such experiments includes:

  • A windows computer to run the HP-compressor
  • A High Pressure compressor at 200-2000 bar of Helium or Argon
  • A medium pressure cell at 150 bar Krypton
  • A medium pressure cell at 50 bar of oxygen
  • A fume-hood to handle samples under gas atmospheres
  • The necessary crygenic tools and deware to handle, cryo-preserve and store samples
  • A micriscope and a lab bench to prepare samples
  • A 4C fridge to store solutions and chemicals

Contacts

for any questions for applications please contact either Philippe Carpentier (philippe.carpentier@esrf.fr), David flot (flot@esrf,fr), Peter van der Linden (vanderli@esrf.fr) or Christoph Mueller-Dieckmann (muellerd@esrf.fr)

Applications

The service offered by the high pressure laboratory (HPMX), is best used prior to a regular visit scheduled on any of the ESRF MX beamlines. Please mention in the regular application for a diffration experiment that you also would need to use the high pressure laboratory.  

Please get in contact with us at least two weeks prior to your scheduled trip by writing an email either to flot@esrf.fr or to philippe.carpentier@esrf.fr. We need to know your experiment number (MX number), the protein of interest, the date the experiment is planned and the reason why you would like to use the HPMX facility. All samples dealt with at the HPMX facility need to be described in the A-form and in the experiment declaration form for safety approval prior of the experiment.

Note: Any specific instrument or chemical that you plan to bring to the ESRF strictly requires to be declared and approved to/by the safety group.