The laboratories are well equipped with a wide variety of instrumentation for modern biomedical research - DNA sequencers, FPLC and HPLC systems, spectrophotometers, stop-flow fluorometers, scintillation and gamma counters, phosphoimagers, thermal cyclers and ultracentrifuges.
The answers to major problems in biomedicine and the keys to better treatment of cancer and AIDS are buried in the intricate, three-dimensional shapes of the molecules that make up our bodies. By mapping out the three-dimensional structures of important biological molecules, we can better comprehend how these molecules work.
Wayne State University's Advanced Laboratory of Macromolecular Crystallography is one of only a few laboratories in the world to run a high-powered X-ray generator capable of producing an X-ray beam that is ten times stronger than conventional generators, and is almost as strong as that of a particle accelerator. This high-flux source allows structure determination of microcrystals that could normally be handled only at synchrotron facilities. Furthermore, structure determination of normal size crystals is performed in a fraction of the time. As a partner in the Michigan Life Sciences Corridor, Wayne State is participating in a statewide initiative to advance studies in structural biology. The consortium is currently building a $13 million X-ray diffraction station at the Advanced Photon Source of the Argonne National Laboratory in Chicago-the strongest source of X-rays in the world. The facility will allow WSU researchers to access four different X-ray beamlines associated with two undulators and two bending magnets. The high-flux X-ray generator and other facilities associated with ALMC are available for partnership ventures between Wayne State investigators and Michigan companies seeking to pursue structural studies of biomedical interest.
The ALMC is located in Gordon Scott Hall, WSU School of Medicine. The major equipment and intellectual environment at WSU is shared with the macromolecular crystallography groups led by Dr. Brian Edwards, Dr. Domenico Gatti and Dr. Ladislau Kovari. The ALMC facility is managed by Dr. Philip Martin. Our students and postdoctoral fellows mix freely in our adjoining space, giving a wide intellectual experience where we can all help each other.
The X-ray diffraction equipment consists of a Rigaku FR-D high brilliance rotating anode generator, an RAXIS HTC image plate system mounted on a two theta stage, a HighRes2 mirror system. Crystal ultra-low temperatures are reached with a Rigaku X-stream 2000 cryogenic system.
Leica StereoZoom microscopes are used to examine crystal growth and to take photographs of crystals. A DynaPro MSXTC/12 dynamic light scattering instrument is used to measure mono- and poly-disperse nature of the protein samples prior to performing crystallization trials.
Nuclear Magnetic Resonance (NMR) spectroscopy provides another method for determining the structures of biomolecules. However, since the biomolecules are in solution, one can also obtain information regarding dynamic regions of proteins, structural and kinetic information about how protein binding partners come together and quickly scan structural rearrangements of enzymes as they perform catalysis in real time. Since the majority of the atoms in a protein can be labeled with spin active (NMR observable) nuclei, one can also probe the chemical properties of atoms in a biomolecule using this technique.
The Department of Biochemistry and Molecular Biology has state-of-the-art NMR instrumentation that graduate students have complete access to. Spectrometers available for student use on the Wayne Campus include 700 MHz, 600 MHz, 500 MHz and 400 MHz spectrometers. In addition, the department participates in a state-funded program that allowed us to recently purchase a 900 MHz spectrometer, which will be located close to our university and available for our students to use.
Numerous dedicated computers for scientific instrumentation are scattered throughout the research areas. All systems are linked to the Internet. The Computing and Information Technologies (C&IT) resource of WSU is an especially versatile and user-oriented facility emphasizing interactivity. The Medical School Information Systems (MSIS) is comprised of four departmental units: Network Operations, Desktop Operations, and Systems Development. In addition to providing support services to students, faculty and staff at the School of Medicine, MSIS is continually exploring new technologies to ensure that the SOM computing experience meets the growing and changing needs of today’s academic environment. High performance computing resources located in the Department of Biochemistry and Molecular Biology include Silicon Graphics UNIX and Linux workstations. These computer workstations are used for research in structural biology. The suites of crystallography programs available are: D*TREK, HKL2000 and X-gen for data reduction, CCP4, CNSSolve, PHASES for structure determination and refinement, XtalView, QUANTA for molecular graphics, MODELER for homology modeling and DOCK for ligand docking and design.
Cell Lines Resource
The Karmanos Cancer Institute has developed a number of unique cell line resources. In particular, a number of lines are maintained and distributed by the investigators in the Breast Cancer Program with special expertise in their development. The MCF-7 breast cancer line is a major research resource generated from this facility. A more recent resource is a set of immortalized lines, designated MCF-10 and MCF-12 derivatives, that provide models of nonmalignant human breast epithelium. One of these derivatives, MCF-10AneoT, is of special interest for studies on the development of human breast cancer. Animal and Human Tumor Resource The Animal and Human Tumor Resource facility is responsible for the acquisition and distribution of tumor cell culture and xenograft models most commonly used by investigators in chemotherapeutic, biochemical and biological studies. Quality control of tumor stocks includes MAP testing for viral contamination; determination of bacterial contamination; histology; and biologic behavior. In addition, staff of the faculty provide protocol design assistance, demonstration of techniques, data analysis methodology, and technical assistance as required.
Confocal Imaging Core Facility
The Confocal Imaging Core Facility includes a Zeiss LSM 310 microscope and associated Silicon Graphics workstation. With recent upgrades to the microscope and computer, the Confocal Imaging Core Facility offers state-of-the-art technology for the evaluation of up to three fluorophores in a single specimen. Two lasers can be used for simultaneous analysis and the uv laser can be used for an additional analysis within seconds of the lasers. Both fixed and live specimens can be analyzed, with the live specimens able to be analyzed at temperatures ranging from 15°C to 65°C. Optical slices of images can be reconstructed in three dimensions on the Silicon Graphics workstation. The image recording systems allow one to obtain publication quality prints, as well as slides, from either the Zeiss Monitor or the Silicon Graphics monitor. Therefore, images for publications, grant applications and presentations are available immediately without having to go to outside image processing facilities. An important advantage of the Zeiss LSM 310, in comparison to other confocal microscopes, is its user friendliness. After initial training, individual investigators are capable of operating the microscope with minimal technical assistance.
Pharmacokinetics and Metabolism Core Facility
The Pharmacokinetics and Metabolism Core Facility provides expertise and equipment to perform pharmacokinetic studies as well as studies on drug metabolism in vivo and in vitro. In addition to the capability to perform studies on plasma pharmacokinetics in either human or murine models, the kinetics of anti-cancer drugs can be monitored in tumors and major organs in the murine models. The facility also has the capability to analyze drugs and metabolites in urinary and fecal excretions and to measure the amount of free drug in the serum. Cellular pharmacokinetics and assessment of drug uptake into tumor cells and in vitro studies on tissue culture lines are also available. In addition, the facility has the capability of analyzing drug metabolism in tumors in vitro, in tissue culture lines of sensitive tumors, in tissue homogenates and subcellular fractions such as microsomes or even purified enzymes. Methods of analyses which are routinely available through the facility include high performance liquid chromatography (HPLC) and gas chromatography (GC). The capability of this core is currently being enhanced by a collaboration with the Institute for Chemical Toxicology, which maintains a series of cell lines that have been genetically engineered to express specific cytochrome P450 cDNAs. These lines allow identification of the specific enzymes that contribute to the metabolic disposition of a particular compound. Both cytotoxic and genotoxic assays are performed routinely.
Quantitative Image Processing Systems Core Facility
A quantitative image processing system (QUIPS) for fluorescence microscopy consists of an epifluorescence microscope interfaced to a computer equipped with appropriate software to allow for the acquisition and quantitative analysis of multicolor fluorescence images. This system allows performance of comparative genomic hybridization (CGH), physical mapping of probes (Flpter analysis), and various other techniques including metaphase finding for translocation analysis and cell-based phenotype/genotype analyses. This system is useful to any investigator interested in using the tools of modern molecular cytogenetics as they apply to many disease processes.
Molecular Genetics Core Facility
The new Molecular Genetics Core Facility will serve two primary functions: (1) The core will provide molecular genetic services (DNA sequencing, genomic and cDNA library screening, mutation detection, microarray applications including bioinformatics and analysis); and (2) will provide free-of-charge consultative services. The core will also provide training opportunities in specific molecular techniques to interested researchers on a fee-for-service basis.