Instruments

The protein crystallization platform has multiple robotic devices:

  • A solution-handling robot mixes the crystallization chemicals from stock solutions and prepares the crystallization plate.
  • A nanodispensing robot sets up the crystallization drops by mixing the protein with an aliquot of the well solution.
  • A robotic plate mover presents the plates to the microscope according to their imaging schedule, and digital images of each drop are stored in the database.
  • The Hamilton Microlab STAR robot mixes the crystallisation chemicals from as many as 60 stock solutions at a time. The robot can accurately handle many types of liquids from alcohols to viscous PEG solutions, down to volumes as low as 15 µl. All crystallisation chemicals are first pipetted into a deep well block at 0.5 - 1 ml volume, mixed and then aliquoted to the crystallisation plate.
  • SPT Labtech’s mosquito LCP® liquid handling technology provides precise and repeatable nanolitre pipetting, every time, irrespective of liquid viscosity or environmental conditions. It can pipette accurately down to 50 nl volumes (volume range of 25 nL to 1.2 μL), but we prefer to use a minimum of 100 nl of protein, and 100 nl of well solution. During the drop dispense step, the plate is covered to minimize evaporation. Immediately after the setup is complete, the plate is sealed with UV-friendly transparent film.
  • The LCP technique for crystallizing membrane proteins can be difficult and time-consuming to set up by hand, as it utilises highly viscous lipid mesophases to reconstitute proteins. SPT LabTech has overcome these problems with mosquito LCP, a dedicated pipetting instrument for automated LCP screening set-up.
  • Using a positive displacement syringe with automated tip positioning, mosquito LCP provides accurate and repeatable dispensing of the LCP drops.
  • The Minstrel DT UV from Rigaku is an ultraviolet and visible crystal imaging and protein crystal monitoring system. This robotic instrument is a major advance over previous visible light microscopy technology because its UV technology can find crystals in complex drops, and easily distinguish protein crystals from non-protein crystals (such as salt).
  • PixRay/IceBear allows users to remotely view and score their images over the web, across platforms such as Mac or PC.
  • The robotic handler feeds plates to the Xtal Focus imager as determined by a preprogrammed schedule. The plate history and all images are stored into the database, and can be remotely checked by users through either PixRay or IceBear interface. 
  • Rigaku Xtal-Lab Synergy S microfocus sealed tube X-ray diffractometer with 0.1 um beam size, adjustable slits, four circle kappa-goniometer and cryo-cooling for data collection
  • For both macromolecular and small molecule data collection (Cu-anode)
  • HyPIX-6000 hybrid pixel array photon counting detector for very fast read out and very low noise
  • Xtal-Check in-situ plate-screening option for screening crystals directly from the crystallization plates
  • Sample optimisation with HPLC-MALLS (Shimadzu LC-10Ai pump, Wyatt Technology miniDAWN TREOS and Optilab REX detectors) - Analysis of sample monodispersity and molecular weight by analytical size exclusion chromatography and static light scattering with microliter volumes.

 

  • Sample optimisation and protein stability measurements with Thermofluor - highly recommended as a first step before crystallization. A pre-screen helps optimizing protein to dye ratios. Using a full buffer and salt screen (96 wells format) we can measure the thermostability of your protein based on fluorescent dye binding.
  • Temperature-gradient screening system for protein crystallization (Thermo-screen)
  • Can be used for intial screening of temperature for crystallization on a gradient PCR machine e.g. from 40-370C on a 96-well plate