Richard Pestell

The Lankenau Institute for Medical Research

Wynnewood, Pennsylvania

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.

Learn More about LIMR: http://www.limr.org/

WEBSITES RELEVANT TO TRANSGENIC STUDIES:

Non-murine transgenics: