Professor Pestell and his researchers have access to facilities at three Institutes, expanding their repertoire of capabilities, including:
1. The Lankenau Institute for Medical Research (LIMR) (14 PI, 41 research personnel),
2. The Blumberg Institute (15 PI, 26 faculty, 205 researchers at the entire Biotechnology Center), and
3. The Wistar Institute (32 PI, 289 researchers).
the Lankenau Institute for Medical Research
Animal: The Lankenau Institute for Medical Research has a Central Animal Facility accredited by AAALAC. The transgenic and knockout mice used in this project have been generated and well characterized in the preliminary studies.
The Proteomics Facility used for the preliminary studies is located at the Wistar Institute and accessible by user fees. This Facility consists of multiple sections, for protein sequencing, amino acid analysis, protein/peptide purification by HPLC or electrophoretic techniques (e.g. electro blotting), peptide synthesis, and mass spectrometry. Other facilities are located at the Lankenau Institute for Medical Research (LIMR) and include Bio imaging, Biostatistics, Cytogenetics, Flow Cytometry, Pathology, and X-Ray Crystallography and other facilities located at the Wistar Institute. The services of the facilities are provided on a charge-back basis. All the LIMR facilities are located on the same floor or within two floors from Dr. Pestell's laboratory in the same building. The administrative office for LIMR is located on the floor below the PI’s office. There is a conference room and library that contains key journals. The transgenic mice are housed on the same floor as the laboratory providing easy access for the proposed experiments. The laboratories are well equipped for molecular biology experiments. The investigators of LIMR, the Blumberg Institute and the Wistar Institute, are focused on cancer research and represent a substantial collective group of approximately funded laboratories. Many laboratories are actively involved in studies of transcriptional control, innovative transgenic mouse modeling and translational research initiatives. This Institution provides a rich research environment that fosters the exchange of ideas and provides a number of relevant seminars, symposia, and journal clubs.
LIMR (affiliated with Thomas Jefferson University) is a state-of-the-art research facility located in a modern, 53,000-square-foot three-story building contiguous with the Lankenau Medical Center, which contains a 320-bed tertiary care teaching hospital in Wynnewood, PA. LIMR and the Medical Center have a vibrant Infectious Disease group. All equipment required for the project is available at LIMR.
Laboratories: Within LIMR are sixteen 1,137-square-foot laboratories located on the perimeter of the first and second floors that are designed and equipped to support research utilizing the latest molecular and biochemical techniques. All laboratories are equipped with at least one fume hood and one to two laminar flow hoods serviced with vacuum and gas. CO2 incubators are supplied by a central CO2 generating system located in the loading dock adjacent to the ground floor. All the laboratories are well equipped with basic equipment necessary to perform molecular and biochemical techniques, including micro-centrifuges, balances, shakers, water baths, SDS PAGE equipment, transblot apparatus, agarose gel electrophoresis equipment, PCR machines, real time qPCR, power supplies, gel dryers, pH meters, warm/stir plates, vortexes, a variety of pipeting devices, lyophilizers, rotoevaporators, ultrafiltration apparatus, 4 hybridization ovens, deli-style refrigerators, microwaves, ELISA plate readers, and spectroflurometers. Low temperature freezers are in a central location as are liquid nitrogen storage units supported from a central tank.
Major Equipment: LIMR has all the equipments required to run the proposed project. The Institute has an assortment of microscopes for performing dark phase and light field and fluorescent microscopy, including 3 Wild M3Z dissecting microscopes, a Leitz Labovert inverted compound microscope and a Nikon Diaphot inverted compound microscope, which are both equipped with micromanipulators, Axioplan and IMT-2 photomicroscopes, a Zeiss Axioplan Phase Contrast Fluorescent microscope, a Zeiss Axioscop 20 Fluorescent microscope equipped with a Optronics DEI 750 video camera and a Video Sony UP-D 5600MD printer, an Olympus BX60 fluorescent/brightfield microscope dedicated for image analysis, an Olympus BH2 fluorescent/bright field microscope with fully automated Prior stage for cell counting, a Canon BX60 fluorescent microscope, a Nikon SMZ1500 fluorescent dissecting microscope and Nikon Confocal microscope. Available on a shared basis are a Varian HPLC system equipped with a IN/US bRAM Model 2 flow-through detector, Vickers M-85 scanning densitometer, Speed-Vac, Bio-Rad Gene Pulser with capacitance extender for electroporation of eukaryotic cells, a BioMag Magnetic separator for magnetic cell sorting, a Turner Model 20e luminometer, and 2 Coulter counters. In addition to the basic laboratory equipment, a central service laboratory has a BD FACSCanto II flow cytometer containing 488nm solid state and 633nm HeNe lasers with 6-color capability, two Cruachem oligonucleotide synthesizers, two ABI Prism 310 Genetic Analyzer for automatic DNA sequence analysis, 3 Bio-Rad DNA sequencing apparatus, ABI Prism 7700 Sequence Detector, and a Molecular Dynamics Phosphoimager/ Densitometer. The Institute also has Biotek ELISA readers. The instruments are available to the entire scientific staff.
Spacious scientific support areas (1005 ft2) are designed into the central core area on each floor: The central glasswash area equipped with three automatic washers, a Getinge/Castle Biohoe autoclave, and two large capacity forced air ovens is located in the core of the ground floor. Installed in the central glasswash is a constant re-circulating high purity reverse osmosis water system (Culligan) that supplies 18 megohm water and steam to the washers and autoclave, respectively. A full-time glasswash technician is employed by the Institute to operate this facility. Common rooms for equipment shared by the entire scientific staff are located on the first and second floor central core areas where there are 4 Beckman Optima ultra-high speed centrifuges, 4 Sorval plus 1 Beckman high speed centrifuges, 3 Packard liquid scintillation counters, 1 Packard Cobra gamma counter, 2 Virtis freeze dryers, 2 Scotsman ice machines, a Gilford 2600 recording spectrometer, a Beckman DU 7500 UV/visible spectrophotometer, 2 floor mounted environmental shakers, a Branson ultrasonic cell disrupter, and in each common service area, a Culligan recirculating water system that supplies the laboratories with high purity water as well as the source of high purity steam for the Getinge/Castle Biohoe autoclave on each floor. A refrigerator/freezer room adjoins the common equipment room on each floor. On each of these floors is a dark room equipped with a copy stand, UV light box, and a film processor (Kodak Xomat in one of the dark rooms and a Konica XRX in the other) and a BioRad Chemidoc for gel and blot documentation and analysis. In addition, a walk-in cold room and warm room are located adjacent to the common equipment room on both of these floors. A Histology Laboratory, Liquid Nitrogen Freezer Room, mechanical space and a Storage Room are located on the perimeter of the Ground floor. A fully equipped histology laboratory is equipped with a RMC tissue processor, 2 Leica/Reichert microtomes and 1 Bright Instruments Cryo-microtome.
In the Liquid Nitrogen Freezer Room, five liquid nitrogen freezers are attached to a manifold system specially designed to monitor and maintain a safe level of liquid nitrogen in each freezer. In addition to liquid nitrogen freezers, this room can accommodate -200C and -800C freezers. A large liquid nitrogen tank is located exterior to the Freezer Room as the source. Also located on the ground floor is a room specifically designed to safely operate a J. L. Shepherd Mark I, Model 68A Cesium Irradiator.
LIMR has a number of Cores in place to support the research programs of all investigators. The Cores include a Histology Core, which prepares and sections frozen and paraffin embedded tissues,; an Imaging Core to analyze microscopic specimens; and a Transgenic Mouse Core, which generates mutant mice. LIMR investigators also have access to the core facilities at Thomas Jefferson University, Wistar Institute and University of Pennsylvania.
Baruch S. Blumberg Institute
The Baruch S Blumberg Institute “owns” and operates the Pennsylvania Biotechnology Center, a 110,000 ft2 research and instructional “incubator” for life sciences start up companies. The Center contains all of the equipment and infrastructure for state of the art molecular biology, drug discovery, and life sciences research, including NMR, DNA sequencing, protein peptide identification, flow cytometry. Faculty of the Blumberg institute have access to these resources either by “right” (as members of the Blumberg Institute) or through vendor relationships (where Center companies provide fee for services access. In addition, of course, Blumberg scientists have access for animal resources, genomics, arraying, DNA sequencing, and numerous other services, by way of institutional affiliations (with, for example, The Commonwealth Medical School), and institutionally arranged relationships with commercial vendors.
For its exclusive use, The Baruch S Blumberg Institute reserves ~16,000 ft2 of laboratory, office and instructional space at The Center, as described below, here:
Laboratory: The labs are modern, “molecular” and “tissue culture” biology oriented labs, remodeled in 2007, with laminar flow hoods, chemical fume hood, incubators for culture, inverted (Nikon) and direct (Olympus) microscopes, access to direct and inverted imaging microscopes, including a Leitz De-convolution microscope; FACs (FACs Caliber, and FACs Arias), and several mass spectrometry and one NMR system. There is equipment for ELISA (plate washer-reader system) and glycan assays (Dionex HPLC system) and development, which are within our lab, and immediate access to a ThermoFinnigan LTQ.
Institutional environment: We are located at The Pennsylvania Biotechnology Center of Bucks County (PA Biotech Center), in which we are co-located in a 3 building, 110,000 sq ft complex with The Hepatitis B Foundation and Baruch S. Blumberg Institute as well as 40 small start-up companies. There is thus a concentration of drug discovery oriented scientists, here, along with a strong “commercialization” theme. There are two companies on-site that provide CRO type contract services, for medicinal chemistry, and flow cytometry.
Computers: Personal and on-line lab equipment computers (MAC and PC) are served by a password protected, secure network linked by 100BaseT and Gig-ethernet and secure WiFi, with central network storage. There are several networked computers handling research needs in the PI’s laboratory and offices. Several printers, plotters, scanners and computer-linked instruments are located throughout the laboratories. A full-time IT Manager oversees the hardware, software, with over 20 years of experience.
Office: The PI has a private office with a personal computer (MAC), a printer and shared offices for his research staff, each whom has their own computers and shared printer. There is a centralized administrative office located adjacent to the laboratories and research offices. There is also access to 1 large lecture hall and several classrooms and conference rooms (within Blumberg Institute) as well as small library and tele- and video-conferencing equipment for meetings, lectures and classes.
Administrative Support: A central administrative office is staffed with a full-time Office Manager, a Grants Administrator equipped to handle all matters relating to this project, including compliance, budgeting, and staffing as well as a Scientific Facilities Manager equipped to handle all regulatory and compliance matters related to the scientific laboratories and equipment. The admin office has a fax machine, scanner, color printer, and coordinates all shipping and receiving for the entire building, along with full mail services.
Drug Discovery An unusual feature of the Blumberg is its resources for therapeutic drug discovery and development. In addition to its own diverse compound library of small molecules (~120,000), and its natural products collection (one of the largest and most diverse in the world, the result of a gift of the entire US Collection of Merck and Schering Plough’s collection (representing ~70% of all plant, bacteria and fungi species on earth), the Center has an “industry standard” robotics and bioinformatics screening unit, managed by a former Director of High Through Put Screening, from Johnson and Johnson. Biomek and EPIC robot systems for screening, and individuals with significant drug discovery and development experience are on site to collaborate and assist an investigators’ research.
Shared Facilities/Major Equipment
(Relevant to the project, and accessible):
Autoclaves, glassware/kitchens/ tissue culture rooms, dark rooms, microscope dark rooms and Nikon digital microscopy, with IF attachments; liquid scintilators, FACS caliber, Guava, LTQ Mass Spec (with on line HPLC); ABI QTrap, multiple HPLC systems, FPLC, ABI array printer, confocal microscope, NMR, Biomek 200 Robot and Epic high throughput sequencing systems and a FACS Aria (via Flowmetric, on site) are all housed within the same building and are available within a 1 minute walk from the PI’s laboratory BSL-3 work, but currently used as BSL-2+), equipped with a biosafety cabinet and two CO2 incubators, a microscope, a refrigerator, -20 and -80 freezers.
Waters Alliance 2690 HPLC systems with enhanced and Fluorescent detectors; ABI Voyager DE Pro MALDI-TOF Mass Spectrometer; 1 Agilent LS Mass Spec (LCMS6120) ThermoFinnigan LTQ linear ion trap Mass Spec; ThermoFinnigan LCQ Mass Spec w/ a splitter ABI Q-trap Mass Spectrometer; LI-COR Odyssey Imaging systems for detection of proteins.
Real Time PCR Systems – (Applied Biosystems-7000 Prism,7500 Fast, Roche-LightCycler LC-480, LightCycler II). Olympus Fluorescent Microscopes (models BX60, IX70, IX81)
Nikon Deconvolution Fluorescent Microscope – Eclipse TE 2000-U
Guava Flow Cytometer – EasyCyte Plus; 1 Becton Dickinson Flow Cytometer – FACSCalibur
Gel Imaging systems (Alpha Innotech-Chem Imager, Kodak-Gel Logic 1500, Bio Rad- Molecular Imager FX (2); Hope Film Processor – Micro Max; Waters HPLC systems for protein purification.; Liquid Handling Robot – Beckman Coulter – BioMek NX MC 2 Liquid Nitrogen Freezers – MVE – 1500 Series-19; Liquid Scintillation Counter – Wallac – 1409 DSA 2 Plate Readers - BioTek-Synergy2, AID-EliSpot
Plate Readers(Scintillation and Luminescence)–Wallac-1450 Microbeta Trilux, Perkin Elmer- Sonicators – Misonix-XL2020;
Spectophotometers – Thermo Scientific – NanoDrop 1000, NanoDrop Speed CentrifugesSorvallRC5B; 2 Ultra Centrifuges – Beckman – L8-70, L8-70M Thermocyclers, Eppendorf-MasterCycler Pro S, MasterCycler Gradient (2), Biometra- Thermocycler: Techne-GeneE Ultra Pure Waters – Milliipore-MilliQ Synthesis 1 300MHZ NMR
The Wistar Institute
The faculty and senior scientific staff of The Wistar Institute have assembled state-of-the-art technologies into Shared Resources that function as engines of discovery for biomedical research initiatives.
Wistar’s scientists share these resources, when possible, with the greater research community. Wistar Shared Resources, also known as core facilities, are available to Wistar investigators, affiliates, and outside researchers. Many of the Shared Resources are supported by a Cancer Center Support Grant (CCSG) awarded by the National Cancer Institute (NCI) to The Wistar Institute Cancer Center.
These Shared Resources include: the Animal Facility, Bioinformatics, Flow Cytometry, Genomics, Imaging, Molecular Screening & Protein Expression, Proteomics and Metabolomics, and a Biomedical Research Support Core. Additional Shared Resources include the BSL-3 Facility, Histotechnology and a Research Supply Center.
All researchers are encouraged to contact a Shared Resource Director to discuss available services. Contact us and find more in-depth information about Wistar’s Shared Resources: Learn More
Animal Facility: The Animal Facility facilitates research through humane and efficient management of animal populations. The vivarium operates as a modified barrier facility and is equipped with quarantine and a procedure room, holding rooms with biosafety cabinets, an imaging/holding room equipped with a PerkinElmer IVIS SpectrumCT imaging system, and additional support areas. Wistar’s Animal Care and Use Program, overseen by The Wistar Institutional Animal Care and Use Committee, is fully accredited by AAALAC International since 1998, has an assurance on file with the Office of Laboratory Animal Welfare at the NIH, and is a registered USDA research institution.
Bioinformatics: The Bioinformatics Shared Resource continuously develops new and efficient approaches to data analysis as a response to emerging research needs. Facility functions include: statistical analyses and computational modeling for all types of high-throughput data, advanced bioinformatics tools for integrative cancer biology, and data management.
Biomedical Research Support: The Biomedical Research Support Core (BRSC) provides a robust infrastructure to support mechanistic, patient-oriented research. The BRSC manages the resource commitment associated with clinical studies, including supporting compliance with regulatory directives governing research in Human Subjects. Services include phlebotomy, tissue microarrays, collection of pathological specimens and support for clinical studies. This includes data collection, storage and extraction, data quality control, site monitoring, regulatory reporting, and connection with statistical teams for data analysis.
Flow Cytometry: The Flow Cytometry Shared Resource provides users with the technological resources and professional assistance for high quality, multiparameter flow cytometry analyses and sorting. Facility personnel aid investigators in creating efficient and cost-effective experimental designs, through optimizing cytometry-specific reagent and fluorochrome selection, and offer assistance in operation of analysis instruments. Technical support is also provided for analyses of flow and imaging cytometry data for publication, presentation, and inclusion in grant applications, management of cytometric data (storage, archiving, and retrieval), and management of a site license for low-cost analysis software.
Genomics: The Genomics Shared Resource serves as a hub for consultation and scientific interaction relating to nucleic acid-based methods. It provides expertise and support to insure the best possible outcomes for genomic related projects. The Facility supports several state-of-the-art platforms for a wide variety of nucleic acid-based studies, including massively parallel sequencing as well as routine capillary sequencing. Consultation and assistance with experimental design and for the development of custom services are encouraged.
Histotechnology: The Histotechnology Shared Resource provides services for fixing, processing and paraffin or OCT-embedding of all types of tissues for light microscopy (e.g. routine stains, immunohistochemistry or in situ hybridization). The Facility staff performs routine hematoxylin and eosin staining, as well as specialized staining and slide preparation for immunohistochemistry and in situ hybridization. Frozen sectioning is also available, including consultation regarding freezing and fixing techniques to optimize experimental results.
Imaging: The state-of-the-art Imaging Shared Resource provides access to standard and advanced optical imaging systems capable of reaching these goals and offers assistance with advanced image analysis solutions. Researchers may be trained for unassisted use of all core instrumentation, while full service assistance by facility staff is also available for qualitative or quantitative image capture. The Facility also offers expert technical assistance with experimental design to optimize imaging results, enabling users to get more out of the imaging technology.
Molecular Screening & Protein Expression: The Molecular Screening and Protein Expression Shared Resource fosters collaboration by providing expertise in biochemical and cell-based assay development for high-throughput screening and compound profiling. Such assays enable researchers to identify small molecule compounds which interact with a target protein of interest. These compounds can then be used as tools to further study the target proteins function and signaling pathways in cells.
Proteomics & Metabolomics: The Proteomics and Metabolomics Shared Resource provides high sensitivity proteomics and metabolomics analyses using state-of-the-art mass spectrometry instruments and methods. Consultation with facility staff concerning experimental design and sample preparation is recommended prior to sample preparation to ensure optimal experimental design.