USC School of Pharmacy Core Facilities USC School of Pharmacy maintains multiple core laboratories with state-of-the-art equipment suites and appropriate research staff support, providing facilities not typically maintained in individual labs. The three main laboratories, Translational Research Laboratory, Histology and Lentiviral Laboratory, are all located in the John Stauffer Pharmaceutical Sciences Center. Translational Research Laboratory The USC School of Pharmacy Translational Research Laboratory (TRlab) is equipped with a wide variety of technologically advanced instruments essential for cutting edge biomedical discovery and therapeutic development research. The instruments in the TRlab are available for use by all USC researchers, and training and technical expertise is provided as needed. The laboratories are located on the 4th – 6th floors in the Pharmaceutical Sciences Center. Translational Research Laboratory Equipment Instrument (Manufacturer) Applications Molecular Biology OpenArray Real-Time PCR Performs and analyzes digital PCR experiments for accurate and System (ABI) sensitive quantitation of nucleic acid targets 7900HT Fast Real-Time PCR Performs and analyzes quantitative PCR experiments for accurate System (ABI) and sensitive quantitation of nucleic acid targets Experion Automated Microfludic chip-based electrophoresis analysis of DNA, RNA and Electrophoresis System (Bio-Rad) protein size, quantity and quality. Non-denaturing front-end sample fractionation, comprising a stepEdge 200 Separation System wise extraction of a particle mixture using extracting media of (Prospect Biosystems) increasing densities. ChemiDoc XRS+ System (BioImaging of a wide array of samples from large handcast Rad) polyacrylamide gels to small agarose gels and various blots. Detection methods include fluorescence, colorimetry, densitometry, VersaDoc Imaging System (Biochemiluminescence and chemifluorescence. Rad) Cell & Immunobiology Gene Pulser Xcell Electroporation Cuvette-based electroporation of both mammalian and bacteria and System (Bio-Rad) fungal cell types. Gene Pulser MXcell Microplate-based electroporation of mammalian cells – especially Electroporation System (Bio-Rad) useful for primary and hard-to-transfect cell types. Immunophenotyping, cell cycle analysis, apoptosis, cell proliferation LSR II Flow Cytometer (BD) assays Captures microscopic images from a 96-well plate and automated Pathway 435 High-Content image and data analysis. An additional module allows for multiBioimager (BD) parameter analysis of neurite outgrowth in a high-throughput manner. XF24 Extracellular Flux Analyzer Simultaneous measurement of two major energy-yielding pathways (Seahorse) – anaerobic glycolysis and aerobic respiration in cell cultures or isolated mitochondria in a 24-well plate format. Both magnetic and polystyrene bead-based multiplex analysis of up Bio-Plex Suspension Array System to 100 biomolecules (proteins, peptides or nucleic acids) in a single (Bio-Rad) sample. Variable-mode imager that produces digital images of radioactive, Typhoon 8600 (GE Healthcare) fluorescent, or chemiluminescent samples. Protein/Nanoparticle Characterization DynaPro Plate Reader (Wyatt) Measurement of the hydrodynamic radius of particles, polymers, or proteins in solution in the size range of 2-1000 nm in diameter. Differential Scanning Calorimeter Provides information about protein subunit composition, folding and 8500 (Perkin Elmer) thermal stability; measures binding thermodynamics J-815 Circular Dichroism Provides information on protein secondary structure (alpha-helix, specropolarimeter (Jasco) beta-sheet or random coil); measures thermal denaturation Analytical Instrumentation Jobin Yvon Fluorolog-3 Spectrofluorometer (Horiba) GENios Pro Multifunction Microplate Reader with Injectors (Tecan) F-2500 Fluorescence Spectrophotometer (Hitachi) SpectraMax Gemini EM Microplate Spectrofluorometer (Molecular Devices) Lmax Microplate Luminometer with Injectors (Molecular Devices) 1450 MicroBeta TriLux Liquid Scintillation and Luminescence Counter (Perkin Elmer) EnVision 2103 Multilabel Reader (Perkin Elmer) Steady-state and time-resolved characterization of fluorescent samples Variety of multidetection features including fluorescence intensity and polarization, luminescence and absorbance-based applications. Cuvette-based fluorescence analysis with a 220 to 730 nm wavelength range, scan speeds of up to 3000 nm/min and resolution of 2 nm. Variety of fluorescence-based applications, including top and bottom reading optics for both solution and cell-based assays, dual monochromators enabling selection of any wavelength in 1 nm increments, wavelength scanning, well scanning and auto PMT gain. Variety of luminescence-based applications. Additional injectors allow for dynamic assays. High efficiency, low background counting of beta, gamma, and glow luminescence samples in a 24- or 96-well microplate or a 4ml microvial. Variety of applications based on technologies including fluorescence, time-resolved fluorescence, fluorescence polarization, luminescence, absorbance, and Alphascreen. Histology Laboratory The USC School of Pharmacy Histology Laboratory, located on the 4th floor of the Pharmaceutical Sciences Center, provides services on tissue processing, sectioning and staining. Expert advice is available concerning staining techniques and the interpretation of microscopic images. The laboratory also houses several instruments available for use by USC researchers, and training for their use is provided as needed. Histology Laboratory Equipment Instrument Applications Microm Rotary Microtome Cuts thin sections of paraffin embedded tissues. Microm Cryostat HM 525 Frozen tissue samples are thin sectioned with this instrument. Microm Spin Tissue Processor Automatically dehydrates and paraffin infiltrates tissue samples. STP 120 Microm Tissue Embedding Center Paraffin infiltrated tissues are embedded in wax and mounted on EC 350 cassettes with this instrument. Cuts sections of formaldehyde fixed soft tissues. Sections are Vibratome mounted onto microscope slides. Processes bone samples and other tissues that are difficult to CryoJane mount as frozen sections. Slide-mounted specimens are coverslipped and preserved in Shandon Hyperclean Station mounting medium. Examines and photographs microscope slides. Saves data onto Zeiss Axioskop Microscope CDs. Lentiviral Laboratory The USC School of Pharmacy Lentiviral Laboratory provides services for production of small-scale lentiviral stocks for delivery and expression of transgenes or shRNA of interest. The engineered lentiviral constructs are either provided by the end-users or produced in the lab according to the needs of the customers. The lab also helps with stable cell line generation and characterization.