A decision has been made to decommission the X-ray diffraction system (ICARUS) in the Crystallographic X-ray Facility. Luck of funding for the maintenance/running costs and very little usage were being the main reasons for this difficult decision. All X-ray diffraction experiments from now on will be performed at various synchrotrons.
During its six-and-a-half years of service over 2,000 datasets were collected and nearly 18,000 samples were tested. With its help we have utilised the Sulphur-SAD method of phasing on in-house X-ray source and solved over 20 structures using this method. The ICARUS has taught many students how to use X-ray diffraction for protein structure determination, helped many PostDocs to publish their groundbreaking papers, and PIs to get their important grants successfully funded.
It’s been fun.
“So Long, and Thanks for All the Fish”
The imager has been upgraded with a new visible light camera . The imaging of plates is now three times faster (4.5 min vs 13.5 min for 2-drops plates).
The Windows 7 on the control PC has been upgraded to 64-bit and the HDD has been replaced with Samsung 9850 Pro SSD. The database and web servers have now access to all 16GB of installed RAM. This also allowed for the image processor to run with twice amount of threads.
The facility has two additional crystallisation robots – Mosquito and Dragonfly, both by by TTPLabtech Ltd. See Instruments sections for more details. Book the robots on our Electronic Booking System if you want to use them.
The RockMaker and RockMakerWeb have been updated to version 3.2. The major change is new GUI. The in-house Rock Imager user manual has been updated to reflect the changes, see Manuals section for more information. Access to older version of RockMakerWeb is still available, see the “Quick Links” section for the corresponding link.
The Rock Imager (by Formulatrix, Inc) has been upgraded to its maximum capacity of 1000 plates. The controlling computer has been replaced with a more powerful one running Windows 7. The in-house Formulatrix user manual has been updated to reflect the changes in the procedures and new features (see Manuals section for more information).
The sample changer BruNo2 (by Bruker AXS Ltd) has been installed on ICARUS. The sample changer is compatible with the Uni-Pucks and can accept 5 pucks at a time, i.e. 80 samples. The sample changer is capable of screening crystals or performing X-ray diffraction data collection runs in completely automated manner without user intervention.
It is located in room 1-60. Users who wish to use the sample changer must book it using our Electronic Booking System. The training will be provided by Dr Dima Chirgadze.
The Crystallographic X-ray Facility has now taken under its care and support the Oryx6 crystallisation robot by Douglas Instruments Ltd. It is an Automatic Protein Crystallisation system for protein crystallisation and can be used for performing sitting drop crystallisation, ”MMS”Microseed Matrix Screening, 2D optimisation, additive experiments, microbatch-under-oil and multi-variate optimisation.
It is located in room 1-56. Users who wish to use Oryx6 must book it using our Electronic Booking System.
To all users of the Crystallographic X-ray Facility:
The Crystallographic X-ray Facility can now offer a variety of pre-dispensed into 96-wells plates Crystallisation Screens (see the Resources section of the CXF website for the complete and up-to-date list of kits, as well as the availability of stocks).
The Screens are pre-dispensed into 96-wells, 2-drops, MRC-type plates and are in 1-12 format (compatible with Formulatrix Imager/Hotel software).
The cost is only £8.10 per plate (this includes the UV-transparent seal). Please note the price does not include the cost of plates disposal.
The list of currently available Crystallisation Screens:
see the Resources section of the CXF website for the complete and up-to-date list of kits.
If you would like to use this service, please contact Dr Dima Chirgadze (dyc21 at cam.ac.uk) or Mr Simon Quick (sq204 at cam.ac.uk).
The old equipment in the room 1.81 has finally been decommissioned (R.I.P. BCX3). This room will become the Facility’s Crystallisation Robotics Room, and its refurbishment is well under way. The new equipment, i.e. the crystallisation imager (RockImager 500), will be installed on or around March 28, 2011.
Here is a small report highlighting the statistics for the usage of ICARUS and for the X-ray diffraction data collected for the period from 1 November, 2010 to 28 February, 2011.
During this period we had no breakdowns and only had 2 days of maintenance by Bruker engineers (post-installation checks). The ICARUS is still running on its first filament from the time of installation back in October 13, 2010. Since then, the MICROSTAR generator has run at full power (45kV, 60mA) for 80% of the time. The Cobra Cryostream has been running 24/7 since it was turned on October 15, 2010.
The total number of ICARUS operational days for the period from 1 November, 2010 to 28 February, 2011 is 63 days (weekends, holiday and training closures are excluded). There are now 32 users of the Facility that are trained in the usage of ICARUS.
In terms of the X-ray data acquisition, the users have collected 108 complete datasets that resulted in protein structures. The resolution of the datasets ranged between 1.4 and 3.6 Angstrom, and dataset collection time between 10 minutes and 3 days (please note that none of those datasets included data from lysozyme or thaumatin crystals). We also estimate that about 1000 crystals were tested by users for diffraction during this period. The samples from external users run on ICARUS included amyloid fibrils and carbon nano-tubes.
About 95% of all collected datasets were processed (indexed, integrated, scaled and space group determined) using the PROTEUM2 software (Bruker AXS). The users, including novices and experts alike, are generally pleased with the supplied data processing software, commenting on its ease of use during the setup of data collection experiments as well as during the data processing calculations.