Saint Louis University

A shared-use facility that supports expression, purification, and analysis of research proteins by providing an array of instrumentation consultation on strategies for protein production from small to large scale. Available equipment includes shaking incubators, a New Brunswick Celligen fermentor, Avestin Emulsiflex cell disruptor, AKTA Purifier chromatograph, FlexStand hollow fiber concentrator, analytical ultracentrifuge, Beckman-Coulter analytical HPLC, and a Maxwell16 DNA/RNA/protein extraction system. Contact David Wood (dwood12@slu.edu) for more information.

A recent addition to the Protein Core lab is the Shimadzu AXIMA Resonance™ MALDI-QIT-TOF mass spectrometer. The AXIMA Resonance™ is intended to be a workhorse instrument for protein and peptide characterization through its unique combination of MALDI sample introduction with a Quadrupole Ion Trap and high performance reflectron Time of Flight mass analyzer. In addition to peptide and protein analyses and identification, the instrument permits MS/MS spectra of a wide range of analytes, including pharmaceutical compounds, glycans, lipids, and polymers. MS/MS can be performed on singly charged precursor ions as high as 6-7 kDa.

The Resonance™ has been installed and is in testing mode. Training sessions will be scheduled in the near future on sample preparation, data collection, and interpretation. Instrument use charges for SLU researchers are in the process of cost-analysis.

A state-of-the-art X-ray facility is available to researchers in the department for the collection of high-resolution data on macromolecular samples. The facility has a 0.8 kW MM007 Rigaku generator with VHF optics interfaced to an R-Axis IV++ imaging plate system and a 1.2 kW MM007HF Rigaku generator with VHF optics double ported with two R-Axis IV++ imaging plate systems. A Phoenix robotic instrumentation from Art Robbins Instruments is available for the preparation, screening and imaging of crystals.

The Molecular Graphics Facility is equipped with several Mac Pros with 3D graphics cards, SGI Octanes and UNIX Workstations. Available software includes RIBBONS, MIDAS, GRASP, MOLSCRIPT, MODELLER, GROMOS, WHATIF and the complete suite from SBGrid. The programs allow visualization of macromolecules including stereoscopic views, analysis of properties related to the structure of macromolecules, building and refining models of protein and DNA molecules, and molecular dynamics calculations.

State-of-the-art differential scanning and isothermal titration calorimeters are available to researchers in the department to study protein stability and the thermodynamic components of protein-protein, protein-ligand and protein-nucleic acids interactions. These instruments include a MicroCal iTC200 microcalorimeter for isothermal titration studies and a MicroCal VP-DSC for differential scanning calorimetry.

The Metabolomics Core Facility, operated by the Department of Biochemistry and Molecular Biology and the Center for Cardiovascular Research at Saint Louis University School of Medicine, is a University resource located in the Doisy Research Center. The Facility focuses on small molecule identification and quantification by mass spectrometry. We offer fast, high quality specialized analytical services, and we are open to all Saint Louis University investigators. Instrumentation in this Core includes a Thermo Electron Surveyor LC and Quantum Ultra triple quadrupole electrospray ionization mass spectrometer, as well as two HP6890 gas chromatographs with FID detector and HP 5973 MS detector with both electron impact and chemical ionization sources.

The Bioinformatics Facility is equipped with two Linux servers for support of proteomics and genomics applications. The Bioinformatics Facility provides custom database design and programming, support for microarray analysis, genomic sequence assembly, genomic annotation, and script development for automation of protein and DNA sequence searches. Access to a 32 node cluster for computationally intensive work is available through the facility. The available software includes the MASCOT server for peptide mass searches, NCBI BLAST for local database sequence searching, HMMer for protein domain searches and a MySQL database server.

In 1965, Dr. Elliott purchased one of the first commercially available mass spectrometers ever to be imported into the US. James Shoemaker, M.D., Ph.D., founded the Metabolic Screening Lab in 1989 to use gas chromatography-mass spectrometry to diagnose human metabolic disorders. The facility now includes an Agilent 5973 Mass Selective Detector (Mass Spec) interfaced with a 6890 gas chromatograph accessed by a 7683 auto-injector, which is operated by an HP Compaq PC, and an Agilent 5975 inert Mass Selective Detector (Mass Spec) interfaced with a 6890 gas chromatograph accessed by a 7683 auto-injector, which is operated by an HP Compaq PC. The facility also houses a FISONS MD 800 benchtop GC-MS. Two Hewlett-Packard benchtop GC-MS instruments are also available for use by the department.

The department has three "real-time" or kinetic PCR instruments.

• MJ Research DNA Engine Opticon™ 2 Continuous Fluorescence Detection System: This instrument provides investigators with the ability to perform very sensitive, accurate, and reproducible measurements of levels of gene expression. In addition, this instrument can be used in other applications such as measuring viral load, performing allelic discrimination studies, and optimizing PCR conditions.


• Bio-Rad Chromo4 Real-Time 4-color, 96-well PCR system: The Chromo4 is an interchangeable Alpha unit, bringing real-time quantitative PCR to a high-performance DNA Engine platform and allows upgrading to real-time PCR when coupled with an existing DNA Engine thermal cycler. Crosstalk is reduced by bringing fluorophore excitation and detection directly to each well.


• Bio-Rad CFX96 Touch Real-Time 5-color, 96-well PCR Detection System: This six-channel system combines advanced optical technology with precise thermal control to deliver sensitive, reliable detection. Sample and reagent use are minimized using up to 5-target multiplexing with sample volumes as low as 10 µl. Reactions can be optimized in a single run using the thermal gradient feature and all run data can be visualized at once and exported in multiple formats.

Laboratories within the department contain a variety of specialized instrumentation. These include a BioRad CHEF-DRIII pulsed field electrophoresis system for separation of extremely large molecules of DNA (up to 4 megabases), BioRad Biolistic particle delivery system for biolistic transformations of fungal, plant, and animal cells, multiple HPLC, FPLC, and chromatography systems, and an isolated perfused heart lab with Powerlab software for ventricular pressure and EKG monitoring. The department also has dynamic and static light scattering devices for analysis of macromolecular assemblies, standard gel documentation systems, and fluorescent plate readers and spectrophotometers.

A 44,700-sq.-ft. animal care facility located within the Doisy Research Center is managed by the Department of Comparative Medicine. This facility is supervised by three veterinarians and has multiple technical staff who provide training, colony health surveillance, veterinary and technical support to investigators on a cost basis. They provide consultation and collaborative services to assist researchers in project planning and model development.

Flow Cytometry Research Core houses a FACSCallibur flow-cytometer with cell sorter, a Biosciences LSR II, capable of 10-color analyses, and a Biosciences FACSAria, capable of simultaneous sorting of 4 populations. The Flow Cytometry Research Core has two full-time staff and offers discounted rates to SLU researchers.

Animal Imaging Core Facility is located in the Molecular Microbiology and Immunology Department. They offer non-invasive, small animal imaging using an IVIS Spectrum instrument.

Microarray Core Facility is located in the Molecular Microbiology and Immunology Department. They offer full service gene chip hybridization services using Affymetrix microarray technology.

Research Microscopy and Confocal Core, located in the Department of Pathology, provides a wide range of histological and microscopy services on a fee-for-service basis.

Saint Louis University maintains electronics and machine shops that are available to investigators. The electronics shop provides high-quality, in-house service for all standard laboratory equipment.