Molecular and Cell Biology Core
GeneLab - www.biommed.lsu.edu/genelab
GeneLab, founded in 1990, is the historic service unit of BioMMED. GeneLab provides molecular reagents and services, as well as access to state-of-the-art equipment for Louisiana-based researchers. GeneLab has extensive experience in training and developing Louisiana-based researchers and has become well recognized within the LSU system for providing the necessary environment for the development of extramurally-funded investigators. This has been accomplished by providing a multitude of molecular services and training and by continually seeking funding to expand the resources and equipment within the facility. GeneLab currently provides automated DNA sequencing, DNA fragment analysis, recombinant DNA cloning services, and quantitative PCR analysis. GeneLab also provides reagents and supplies at minimal costs to investigators. Training seminars and wet labs are provided year-round to enable researchers to stay current in the rapidly evolving field of molecular technology. Major equipment available for INBRE investigators include: two Applied Biosystems 3130 Genetic Analyzers, four real-time PCR instruments (two ABI 7900HT Sequence Detection Systems, one ABI 7500fast Sequence Detection System, and one Roche LightCycler 480 II), Alpha Innotech Fluorchem Q imaging station, BioRad BioPlex, Agilent 2100 Bioanalyzer, two MJ Reseach PTC-200 Peltier Thermal Cyclers, Bio-Rad GenePulser Xcell Electroporation System, Eppendorf Centrifuge 5417C, Qiagen 4-15C tabletop centrifuge, an Amaxa nucleoporator, Turner Designs TD-20/20 Luminometer, NanoDrop ND-1000 Spectrophotometer, Molecular Devices SpectraMax M2 microplate reader, Accuri C6 Flow Cytometer, and various computers and software and other small equipment necessary for standard molecular biological practices.
Molecular and Immunological Assistance
Molecular biology support for individual projects will be provided by Vladimir Chouljenko (molecular biology, qPCR, BioPlex, Imaging applications). These personnel and other research associates employed by BioMMED will work with individual investigators to help them adopt and perform state-of-the-art methodologies and techniques to clone, and sequence individual genes, to express genes using a variety of prokaryotic and eukaryotic expression systems, to purify proteins for immunological and other uses, to quantify gene expression using the BioPlex instrument and qPCR assays, as well as flow cytometry.
Viral Vector Laboratories (VVL) - www.biommed.lsu.edu/VVL
The establishment of this laboratory was funded by a Louisiana Board of Regents/Louisiana Department of Economic Development Governor’s Biotechnology Initiative Program Grant, which established a cancer treatment modalities research program within BioMMED. However, this laboratory unit has expanded beyond cancer research to provide viral vector and expression systems for numerous investigators and projects. Currently, the VVL generates adenovirus, vaccinia virus, herpesvirus, retrovirus, baculovirus, VSV, and adeno-associated viral vectors for expression of foreign genes and high efficiency delivery to in vivo and in vitro systems. These services and training in viral vector technology will be available to INBRE investigators. In addition, the tissue and cell culture facilities that are associated with this unit can be used to assist and train LBRN investigators in the proper culture and maintenance of cells, as is necessitated by some LBRN projects. The equipment available for LBRN investigator training and cell culture are: 5 BSL2 biosafety containment and tissue culture hoods; 5 double stacked CO2 incubators; 5 -80?C freezers; a large liquid nitrogen cell storage unit; various waterbaths, freezers, and centrifuges for cell and virus preparations.
Protein and Antibody Production and Purification Laboratory (PAPPL) - www.biommed.lsu.edu/PAPPL
The Protein and Antibody Production and Purification Laboratory has recently expanded to occupy substantial space (over 2,000 ft2) in the LSU main campus (Wilson Hall) in close proximity to the LSU Life Sciences Building and the LSU departments of Biology and Chemistry as well as the LSU-based Emerging Technologies Center. PAPPL has established collaboration with physical chemists, crystallographers and rational drug design faculty of the LSU departments of Chemistry (MacNaugtan) and Biological Sciences (Newcomer, Lee). PAPPL produces recombinant protein through bacterial expression, baculovirus expression, in vitro transcription/translation, or mammalian viral or cellular expression systems, depending on an investigator’s application and the protein of choice. Proteins are purified via affinity chromatography, ion exchange chromatography, reverse-phase chromatography, and/or HPLC. Purified protein can then be used directly or can be modified (FITC, HRP, AP, biotinylation, etc.) to suit the investigator’s needs. PAPPL also generates mouse or rabbit monospecific polyclonal antibodies against purified protein immunogens or through direct DNA immunization. PAPPL offers hybridoma production and monoclonal antibody screening, characterization, and purification. The following equipment is available to INBRE investigators: a Biacore X100 biomolecular interaction system, a BioRad Profinia Protein Purification System, an Amersham/Pharmacia fully automated chromatographic AKTAExplorer FPLC system; New Brunswick bacterial fermentor; baculovirus cultivation incubators; large bacterial shaker; and HPLC facilities.
Non-Invasive Whole Animal in vivo Imaging Laboratory (WAIL) - www.biommed.lsu.edu/WAIL
The WAIL has evolved from being solely an imaging facility to a research unit that produces cancer type-specific reagents for monitoring and quantifying tumor progression and metastasis. The unit has developed specific reagents for the development and isolation of multiple promoter-multiple reporter based cell lines and viruses (reporter systems include EGFP, RFP, luciferase, and combinations thereof) that can be imaged within live animal systems. Moreover, the laboratory has expanded to a Division of Laboratory Animal Medicine (DLAM) approved facility with the ability to temporarily house research animals and perform necessary procedures pertinent to imaging protocols. The equipment available within this laboratory to LBRN investigators are: an upgraded Xenogen IVIS 50 whole animal imaging system with fluorescent and chemiluminescent detection abilities; an isofluorane based anesthesia chamber; continuously controlled anesthesia unit for use during imaging; a Kodak In-Vivo FX Pro Imaging System with X-ray capabilities used to image luminescent, fluorescent, radioisotopic and radiographic emissions, a Leica Inverted Fluorescent Microscope and imaging station for generation of cell lines and assessment of reagents prior to inoculation; a BSL2 containment flow hood for live animal work and protection;; DLAM approved animal holding and imaging facilities. Furthermore, Also, WAIL has access to x-ray tomography and other x-ray based clinical equipment housed in the Oncology Program of the LSU School of Veterinary Medicine.
Tissue and Cell Culture Laboratories - www.biommed.lsu.edu
In addition to the tissue culture and virus production facilities housed within the viral vector laboratories of BioMMED, LBRN investigators will have access to the tissue and cell culture laboratory available in BioMMED. The laboratory offers a spacious cell culture clean-room suite with four 5 foot BSL-2 laminar flow hoods, five double-stacked CO2 incubators that can be set at variable temperatures, two large liquid nitrogen cryogenic storage units, four small cryogenic storage units, a fully staffed glassware processing and media preparation facility, and various small equipment. The tissue and cell culture laboratory maintains and provides numerous ATCC cell-lines, and provides media, sera, and cell culture based reagents to interested researchers.
The LSU Flow Cytometry Facility (LSU-FACS) - http://www.vetmed.lsu.edu/facs/
The LSU Flow Cytometry Facility is a core laboratory located on the third floor of the School of Veterinary Medicine building on the campus of Louisiana State University. The facility features a Becton Dickinson FACS Aria dual laser flow cytometer/sorter capable of measuring seven fluorescence emissions as well as 4 way cell sorting capacity. In addition, the facility has a Becton Dickinson Calibur dual laser benchtop flow cytometer capable of measuring four fluorescence emissions as well as a sorting option capable of performing single population sorts. The laboratory also features a Becton Dickinson FACScan benchtop flow cytometer with single laser capability and 3 color emissions. Multi color immunophenotyping, cell cycle analysis, apoptosis studies, and measurements of cellular function are examples of applications performed routinely in this laboratory. The mission of LSU-FACS is to supply state of the art techniques and procedures for phenotypic as well as functional analysis of animal and human cells. LSU-FACS administers, standardizes, and performs daily analytical procedures to support ongoing research projects associated with the COBRE-CEIDR grant. Furthermore, LSU-FACS spends considerable time in adopting and developing novel assays needed by individual investigators. Immunophenotyping, cell cycle analysis, apoptosis studies, and measurements of signal transduction are examples of applications which are performed routinely in this laboratory. LSU-FACS provides analytical capabilities for investigators throughout the Louisiana State University System, including the LSU Agricultural Center, Pennington Biomedical Research Center and Louisiana Emerging Technology Center. LSU-FACS has frequently communicated with the TNPRC FACs personnel about the utilization of new assays.
SVM Microscopy Center - http://www.vetmed.lsu.edu/van/MicroscopyCenter.htm
The SVM Microscopy Center is a core laboratory located on the third floor of the School of Veterinary Medicine. The Microscopy Center is a central service of the LSU School of Veterinary Medicine providing micro-imaging support for research and instruction. The facility houses transmission (Zeiss EM 10-C) and environmental scanning electron (FEI Quanta 200), confocal laser scanning (Leica TCS SP2) microscopes as well as cryofixation, freeze fracture, and metal shadowing facilities. Four computer coupled light microscopes are available to provide a variety of contrasting methods and fluorescence imaging capabilities. The inverted fluorescence microscope has a heated stage with flow-through culture chambers available on demand. Laser micro-dissection-capture (P.A.L.M) System integrated with a Zeiss inverted microscope) is also available in the Center. The Center possesses both software and hardware capabilities in morphometric analysis, image enhancement, image analysis, video microscopy, and both digital and photographic image recording and printing. Two dedicated graphics workstations are available as are a variety of printers (ink jet, laser and dye sublimation) and scanners (flatbed and dedicated transparency). Technical support, consultation, and training are available for all equipment.
Other Core Resources
The MCBC also facilitates access to other Core Laboratories located within the LSU.
The Analytical Systems Laboratory - http://www.vetmed.lsu.edu/van/Analytical%20Systems%20Lab.htm
The Analytical Systems Laboratories are central service, comprehensive analytical laboratories, consisting of The Laboratory for Residue Studies, The Equine Medication Surveillance Laboratory and The Analytical Systems Laboratory. The Laboratory for Residue Studies was established in 1991 and provides instrumentation and expertise for the performance of drug and biological molecule pharmacokinetics, metabolism, tissue distribution and analytical method development. The laboratory is also equipped to conduct complete drug profiling using radiolabeled test materials. The Equine Medication Surveillance Laboratory was established in 1987 and has served as the official laboratory for the Louisiana State Racing Commission since that time. The laboratory screens over 10,000 urine and blood samples per year and has developed sophisticated methodology for screening, detection and confirmation of drugs and their metabolites. The laboratory also conducts illegal drug-use testing for the LSU Department of Athletics and serves as a source of information to the racing industry and the public regarding drug pharmacology, metabolism and clearance. The Analytical Systems Laboratory, established in 1985, houses advanced mass spectrometry equipment that is used to support other laboratories as well as the research of SVM faculty and graduate students. Equipment in these laboratories includes, Fisons Quattro II GC/LC/MS/MS with ApcI and ESP interfaces, one HP 1090 II, low-flow HPLC (1 μl/min), three 1090 HPLCs equipped with UVdiode array, fluorescence, electrochemical and radiomonitor detectors, a tissue oxidizer, one HP 5971 GC/MS system and beta and gamma counters for radiolabel analyses. The laboratory is currently developing full capabilities to conduct low- and high-throughput proteomics analysis and is enhancing their abilities to conduct protein purification via HPLC methodologies.
Mass Spectrometry Core Facility - http://chemistry.lsu.edu/site/Facilities%20&%20Centers/Mass%20Spectrometry%20Facility/item1379.html
The Mass Spectrometry Core Facility within the Department of Chemistry offers a broad range of services with six mass spectrometers which include: a Varian Saturn 2200 GC/MS for gas chromatography separations and electron ionization (EI) or chemical ionization (CI); a Hewlett Packard 5971A GC/MS for routine GC separations and EI mass spectrometry; a Bruker ProFLEX III MALDI-TOF mass spectrometer used for mass analysis of synthetic and bio-polymers as well as mid-mass organics; a double-focusing, high resolution Finnigan MAT 900 with electron, chemical, FAB, and electrospray ionization sources and a PATRIC array detector; a Hitachi MS 8000 ion trap mass spectrometer direct infusion electrospray or atmospheric pressure chemical ionization (APCI); and a new Applied Biosystems QSTAR XL nano-LC-quadrupole/time-offlight mass spectrometer for high resolution MALDI, electrospray, and APCI for proteomics work and exact mass measurements. The GC/MSs and MALDI-TOF MS are open-access instruments. The facility is staffed by two expert Ph.D. mass spectrometrists.
LSU Protein Facility - http://science.lsu.edu/explore.cfm/proteinfacility/
The LSU Protein Facility is a jointly supported core laboratory of the College of Basic Sciences and the LSU Agricultural Center and is equipped to synthesize, purify, and characterize peptides and proteins. Specialized instrumentation includes an Applied Biosystems Pioneer peptide synthesizer (Fmoc chemistry), analytical and preparative scale Waters HPLC systems, a Dionex AAADirectamino acid analyzer, an Aviv circular dichroism spectrophotometer to measure spectra of peptides and proteins to determine secondary structure in solution, and a Beckman Optima XL-A analytical ultracentrifuge to determine molecular weights and aggregation states of peptides and proteins in solution.
LSU - School of Veterinary Medicine Library - http://www.vetmed.lsu.edu/library/
The MCBC provides access to the LSU-SVM Library for all LBRN-funded junior investigators by arranging adjunct faculty status in the Department of Pathobiological Sciences. LBRN junior investigators have direct access to both physical and electronic resources of the Library. Located on the first floor of the Veterinary Medicine Building, the Library occupies 7,400 square feet with a seating capacity of 112. Current holdings include approximately 47,000 volumes and 603 current periodical titles, dealing with all aspects of veterinary medicine, as well as selected materials on human medicine, comparative medicine, public health, the animal sciences and other related areas. The SVM Library offers online journal access to hundreds of scientific journals not available to investigators on PUI campuses. In addition, the Library in conjunction with BioMMED offers 3 Current Protocols titles for electronic use through the Library website. These titles are “Current Protocols in Molecular Biology”, “Current Protocols in Cell Biology”, and “Current Protocols in Immunology”. Coordination of these efforts is handled by Dr. Gus Kousoulas in conjunction with Christine Mitchell, SVM Library Director.
Other COBRE Core Facilities
The MCBC facilitates the use and access of the centralized facilities of three other COBREs for all LBRN junior investigators. Of particular interest is the advanced Molecular Immunology and Cell Analysis Core of the LSUHSC-NO COBRE (Ochoa) http://www.lcrc.info/research/cellAnalysis.html, the Bioinformatics Core of the LSUHSC-S COBRE (O’Callaghan) http://www.lsuhscmicrobiology.com/biocore.htm, which has advanced capabilities for microarray analyses including statistical and high throughput analyses, experimental design, data standardization and normalization, data pathway and other confirmatory analyses, data storage, sharing and management and integration of the project-related data into public databases. It is worth noting that this COBRE bioinformatics Core is now co-managed by investigators Urska Cvek and Marjan Trutschl from LSU-S who were previously funded by the INBRE. Finally, the PBRC COBRE (Gettys) Cell Biology and Imaging Core http://labs.pbrc.edu/cellbiology/ has advanced facilities for live imaging of small animals including monitoring of metabolic parameters, etc. PBRC has also transgenic facilities and advanced proteomic facilities that can be accessed through the PBRC COBRE.
Last Updated: Thursday, November 10 2011 @ 11:19 AM CST|Hits: 3,700