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The MX core offers facilities and expertise for growing diffraction-quality single crystals of proteins, nucleic acids, small molecules, and complexes thereof. For all crystallization services a highly pure, monodisperse wet sample (protein, nucleic acid, etc.) or dry sample (small molecule) is required. The facility provides all the other necessary components and tools to conduct the experiments. The Protein Expression and Purification Core is co-localized in the same building and can assist you in generating a crystallography grade sample for structural biology studies on biological macromolecules.


High Throughput Automated Crystallization Screening and Monitoring for biological macromolecules

High-Throughput (HT) crystallization trials in a multi-sitting drop 96 well format setup with an Oryx4 liquid handler

  • High-Throughput (HT) crystallization trials in a multi-sitting drop 96 well tray format setup with an Oryx4 (Douglas Instruments Ltd) liquid handler
  • Use as little as 50 nl of sample per trial
  • 1 drop per well – 15 µl of sample per screen of 96 trials (150 nl/trial) for a 1:1 protein-to-crystallant ratio
  • 3 drops per well – 45 ul of sample per screen to accomplish 2:1, 1:1, and 1:2 protein-to-crystallant ratios
  • Each 96 well tray takes about ~15 minutes to set up


Crystal Growth Optimization

Temperature plays a very large role in sample crystallization. Our facility is equipped with numerous incubators to provide users with a lot of experimental options for optimizing crystal growth

  • A walk in 4 Celsius room that is equipped with a Leica Stereo microscope and small work area
  • 3 full sized temperature controlled incubators set permanently at 20, 16, and 12 Celsius.
  • A half sized incubator that is meant for users who wish to freely experiment with temperature

 

UV Fluorescence and White Light Imaging of Crystallization Drops

We have a Jansi UVEX plate hotel with automated microscope that provides dual white light and UV fluorescence imaging of crystallization experiments. It can distinguish between protein and salt crystals, helping you to prioritize the crystal hits more likely to lead to diffracting crystals. It can also help you identify protein crystals hidden in dense precipitates and decide if a crystal of poor morphology is worth using as seed material for second round screening. The UVEX is equipped with a CCD camera to acquire high quality digital images for presentation and publication.

The lab is also equipped with two Leica stereo microscopes, one mounted with a digital camera and polarizing lens. This represents another resource for triaging your crystal hits and acquiring quality images. The camera also provides 30 fps video that you can use in presentations or as a training tool when teaching lab members crystallography.

 

Small molecule crystallization

We have a favored approach involving an initial volley of eight solvent conditions to test for crystallizing your small molecule(s). We start with basic diffusion and slow cooling experiments using those conditions, then branch out as needed.