Using X-ray crystallography we can determine three-dimensional structures of macromolecules at atomic resolution. X-ray crystallography involves the arrangement of molecules in a regular lattice by crystallization, and exposing the crystals to an x-ray beam. The regularly spaced atoms within the array will generate a unique diffraction pattern, which can be used to solve the structure of the molecule of interest. This includes the structures of macromolecules eg. proteins, nucleic acids and larger complexes.
We have a Scorpion liquid handler robot (provided by Jeffrey Lee, LMP) to create and optimized crystallization screens. We have a Gryphon Nanodrop robot that can be used to set up arrays of 50 nanoliter droplets to minimize the amount of protein or complex needed to perform crystallogenesis screens. This robot also has special features allowing for the crystallization of membrane proteins in Lipid cubic phase or bicelles.
We have a RockImager 1000 and a 182 that are automated crystal imagers that can collect UV and visible light images to help identify protein crystals. We also have stereo dissecting microscopes to help loop the crystals and mount them in an x-ray beam to examine diffraction quality.
Crystals are exposed to X-rays to gather diffraction patterns that are used to calculate the position of the atoms within a unit cell of the crystal.
16th Floor MaRS West Tower Room 1645
University of Toronto
Art Robbins Crystal Gryphon LCP Dropsetter
Formulatrix RockImager 182 Crystallization imager
Trevor F. Moraes
MaRS Center, West Tower
661 University Ave., Rm 1613
Medical Sciences Bldg. Location
6th floor MSB, Room 6206
1 King’s College Circle
University of Toronto
Art Robbins Scorpion Screen Builder
Formulatrix UV Rock Imager 1000 – Room Temp
Formulatrix UV Rock Imager 27 – 4oC (MSB 6317)
Nikon FRAP-LCP and Fluorescence microscope (for optimization of LCP conditions and trace-fluorescence labeled crystallization experiments)
Douglas Instruments Oryx 8 crystallization robot (with LCP attachment)
Art Robbins Scorpion Liquid handler
Oliver P. Ernst
MSB, Room 5316A
1 King's College Circle
Local sources of X-rays for crystallography include:
1. Structural Genomics Consortium – 101 College St. MaRS South Tower
2. BioZone – Wallberg Bldg., University of Toronto
3. SickKids Structural & Biophysical Core Facility
4. Privé Lab: PMCRT (MaRS East tower)
Rigaku MicroMax007 with Eiger 1M detector
Formulatrix RI-182 plate hotel and imager
Formulatrix NT8 dropsetter
Contact Doug Kuntz Doug.Kuntz@uhnresearch.ca