Macromolecular Crystallography Facility
X Ray Crystallography Facility
The determination of molecular structure at atomic resolution by X-ray Crystallography, since it’s’ invention in 1913 by Sir Lawrence Bragg, has made a huge contribution to the understanding of the physics and chemistry of materials. The technique is now routinely used to determine atomic structure of complex molecules such as proteins and/or nucleic acids with the aim to understand the structure-function relationships underpinning life.
The Macromolecular Crystallography Core Facility (MCCF), along with ten others, was established to meet the requirement of the FLS research effort and identified as strategic (core) elements to underpin experimental research within the compass of the Life Sciences. This particular Facility is formed by two distinct laboratories located either at the Smith Building or within the MIB.
The MCCF - MIB component
The MIB component comprises a suite of dedicated rooms, housed on the lower ground floor, accessed through LG 027. It provides support in macromolecular crystallisation and (preliminary) diffraction data acquisition to various structural biologists housed within the MIB. A (limited) crystallisation and data collection service is run for those in need of atomic structures but not familiar with the technique.
The MIB based facility component contains a Microfocus rotating anode generator (Bruker MicroStar), providing a high brilliance (2.7kW) source with tuned focussing optics (Montel 200 mirrors) that develop a tightly focussed, highly monochromatic X-ray beam (CuKα, λ=1.514Å). Diffraction data is acquired via CCD technology (X8 Proteum 4K CCD, 135mm diameter) with typical acquisition rates, depending on sample, in the order of 10s. This provides for full 3-dimensional data sets in about 2 hours for most cases. Cryogenic data collection (at ca. 100K) is enabled by a cold stream generator (Oxford Cryosystems Cobra). A Kappa geometry four-circle goniometer furthermore enables efficient coverage of diffraction space. Driven by proprietory graphical interface (Proteum2), preliminary crystal survey and experimental strategy determination is rapid and efficient. Full data collection and processing, which may be run simultaneously, is run from a second PC. Data processing is by the Bruker SAINT software suite: users may request Windows or Linux licensed copies for personal use.
In addition, the MIB facility includes a dedicated laboratory space containing a number of temperature controlled incubators, a Molecular Dimensions Mosquito robot capable of nanolitre dispensing and a microspectrophotometer capable of recording crystal UV-VIS spectra. Crystallisation trials run by the facility require approximately ~100L of highly concentrated, phosphate-free monodisperse sample to screen ~1000 conditions. Crystal screening and initial diffraction data collection can be provided by the facility.
For further details please contact:
Dr Pat Bryant
email: p.k.bryant@manchester.ac.uk
Phone: 0161-275-5090/5658
Mobile: 07748 148159