Structural & Biophysical Core Facility

The Hospital for Sick Children, Toronto, Ontario
What the facility does

Instrument facility specializing in biophysical characterization methods used for investigating biomolecular structure, stability, interactions, dynamics and enzymology.

Area(s) of Expertise

The Structural & Biophysical Core Facility (SBC Facility) is a shared instrument facility specializing in biophysical characterization methods of various macromolecules, such as proteins, lipids, nanoparticles and small molecules. The technologies and expertise available for users include: molecular interaction analysis, multi-mode detection analysis, structural denaturation characterization, large-scale association and nanoparticle analysis, chromatography techniques, microscopy, X-ray scattering analysis, and single protein crystal diffraction analysis. The tools are particularly valuable for studying proteins, including folded soluble and membrane proteins and intrinsically disordered proteins; their interactions with binding partners; and their self-association, such as in phase-separated states. The facility serves scientists from the Research Institute of The Hospital for Sick Children (SickKids), external academic institutions, and industry. The equipment is accessed using one of two options: either pay-per-use fees or all-instrument access yearly membership fees. Users of the facility are supported by full-time personnel that provide instrument and experimental assistance while having the use of a full suite of laboratory support equipment and tools.

Research Services

User pay-per-use and all-instrument access instrument facility, sample submission for analytical instrument analysis, research project laboratory services, protein crystallography and diffraction services

Sectors of Application
  • Agriculture, animal science and food
  • Chemical industries
  • Environmental technologies and related services
  • Fisheries and aquaculture
  • Forestry and forest-based industries
  • Health care and social services
  • Life sciences, pharmaceuticals and medical equipment

Name of specialized lab

Name of equipment in use

Description of function

Molecular interaction analysis

Malvern automated isothermal titration calorimeter (MicroCal Auto-iTC200)

ITC determines the molecular interaction thermodynamics of small molecules and biomolecules, such as proteins, antibodies and nucleic acids to determine KD, stoichiometry, enthalpy and entropy.

Multi-mode detection analysis

Pall FortéBio biolayer inferometer (Octet® RED96)

BLI can be applied to biomolecule quantitative and molecular interaction kinetics of protein-protein and protein-small molecule binding characterization to determine KD, kon and koff rates.

 

Molecular Devices multi-mode hybrid plate reader (SpectraMax i3x)

SpectraMax plate reader uses a monochromator / filter set hybrid system and contains UV-VIS, fluorescence, fluorescein FP, TR-FRET and luminescence modes with the option of a temperature controlled and compressed gas source environment.

 

Photon Technology International multi-cell spectrofluorometer (QuantaMaster 80™)

QuantaMaster 80™ measures fluorescence and fluorescence polarization (FP) and has a Peltier-controlled cuvette jacket, stopped-flow injection system and separate two-syringe titrator.

 

Beckman Coulter lab automation workstation (Biomek NXP)

Biomek NXP liquid handler contains a 96-well pipetting head (P20, P50 and P250 tips) for repetitive liquid transfer tasks.

Structural denaturation characterization

TA Instruments automated differential scanning calorimeter (Auto-Nano DSC)

Auto-DSC determines the thermodynamic parameters of macromolecules such as proteins, lipids and nucleic acids (Tm, enthalpy, entropy, Gibb’s free energy, heat capacity).

 

Unchained Labs static light scattering / fluorescence protein thermal-denaturation platform (UNit)

 

UNit determines protein melting temperature (Tm) and aggregation temperature (Tagg) simultaneously using temperature scanning and isothermal incubation in a miniaturized multi-well cuvette.

 

Jasco spectropolarimeter (J-810 CD)

CD performs protein secondary structure evaluation and stability determination and is capable of fluorescence spectrum measurements.

Large-scale association and nanoparticle analysis

Malvern NanoSight nanoparticle tracking analyzer (NS300 NTA)

NTA determines nanoparticle size distribution and concentration in a liquid suspension using light scattering and fluorescent labels through video capturing and Brownian motion tracking.

 

Wyatt multi-angle static light scattering, dynamic light scattering and refractive index detectors (miniDAWN TREOS, Optilab® T-rEX)

Refrigerated MALS-QELS-RI is coupled to the Bio-HPLC and is used for determining molecular weight, multimeric state, shape and polydispersity of molecules such as proteins, peptides, nanoparticles and polymers.

 

TA Instruments rheometer (Discovery HR-3)

Rheometer measures viscosity, rigidity, shear and strain of liquids and other materials. Contains a Peltier-controlled sample plate with various geometries available.

Microscopy

Zeiss brightfield / fluorescence microscope with Peltier-controlled stage (Axio Observer 7)

The Axio Observer 7 microscope has a Peltier-controlled stage (4°C to 95°C), adaptive focus stabilization, DIC, Axiocam 702 mono CMOS camera with seven LD and immersion objectives. The Colibri 7 light source provides fast excitation wavelength switching (red, yellow, green, cyan, blue, violet, UV). The internal / external fast filter wheel switching includes DAPI, TRITC, CFP, GFP, Texas Red, DsRED, mCherry, Fura-2 calcium imaging, Cy3/Cy5 and CFP/YFP FRET, FITC FP. Has hypoxia chamber with humidity / temperature controls and CO2 / O2 modules and optical sectioning deconvolution and particle tracking analysis software.

Chromatography

Agilent Bio-Inert HPLC (1260 Infinity II Bio-Inert LC)

Refrigerated Bio-Inert HPLC is coupled to the Wyatt MALS-QELS-RI. HPLC contains a quaternary pump, multi-sampler, column compartment, UV-VIS and fluorescence diode array detectors and fraction collector. Several columns available for method development including reverse phase, carbohydrate, IEX, protein and polymer SEC.

 

GE Healthcare Life Sciences FPLC (AKTA Pure)

FPLC can be used in-line with the SAXSpace. Refrigerated standalone instrument contains multi-wavelength detection, conductivity meter, pH meter and fraction collector. Several columns available for method development.

X-ray scattering and single protein crystal diffraction analysis

Anton Paar automated sample changing small and wide angle X-ray scattering (Auto-SAXSpace) with optional in-line SEC-FPLC (AKTA Pure)

SAXSpace determines shape, compactness and size of nanostructures. Structural and dynamic characterization of materials, including nanoparticles, proteins, pharmaceuticals, polymers, surfactants as well as colloidal and biological samples. Sample cell options, including flowcell, pastecell, µ-cell, rotorcell, tubecell, quartz cuvette within a Peltier-controlled sample stage.

 

Bruker single crystal X-ray diffractometer (D8 Venture)

D8 Venture uses a microfocus, TXS rotating anode (Cu-radiation), standard data collection and heavy-atom or sulphur-SAD phasing. In-situ crystal diffraction can also be performed.

 

Fully equipped protein crystallography laboratory with liquid handlers (ARI Gryphon, Douglas Instruments Oryx4, Rigaku Alchemist), microscopes, incubators and crystal hotel & plate imagers (Rigaku CrystalMation DT)