UPDDI Core Facilities and Equipment

Discovery Technology

The UPDDI occupies a total of 7,275 sq. ft. of newly renovated laboratory space on the 9th floor of the Thomas Starzl Biomedical Science Tower (BST). Residing in this space are 5 individual laboratories with facilities to propagate a wide variety of mammalian, insect and yeast cells; a room for protein production and molecular biology procedures; and a chemicals/preparation room dedicated to reagent preparation.

The UPDDI High throughput screening (HTS) and high-content screening (HCS) facility occupies 1,800 sq. ft. and houses imaging, analysis, and an array of robotic liquid handling systems for bulk delivery of reagents or cells (Thermo Scientific Multidrop Combi, Biotek Microflo, and Titertek MAP-C), as well as for compound dilution and transfer to assay or daughter plates (Perkin Elmer Multiprobe, Evolution EP3, and Janus MDT, and the Velocity 11 Bravo). Two BioTek plate washers are available for washing cells in 96- and 384-well plates.

A variety of detection instruments at the UPDDI enable running every kind of assay including: absorbance, luminescence, fluorescence intensity, time-resolved fluorescence, fluorescence polarization, and high-content fluorescent imaging. Plate reader detection systems include: Molecular Devices SpectraMax M5, M5E multi-mode readers, FlexStation 3 kinetic reader, the Perkin Elmer EnVision multilabel reader, and Topcount scintillation and luminescent reader. There are five automated high-content imaging platforms, the GE Healthcare IN Cell 6000 laser based confocal system, the ArrayScan II upgraded to version IV and integrated with a Zymark Twister plate handler, the ArrayScan VTi wide-field system integrated with a Caliper Twister II plate handler, and two Molecular Devices Image Xpress Ultra laser point scanning confocal systems with MetaXpress and AcuityExpress software and integrated with a Thermo CRT Catalyst Express plate handler.

Other instrumentation includes a GE Healthcare Storm Phosphorimager, a Biacore T100, Bio-Rad iQ5 Real-Time PCR system, InVivo2 400 hypoxic workstation, Waters Model 2690 HPLC with a 2487 spectrophotometer, a Waters Model 994 photodiode array detector, a GE Healthcare Aktä FPLC, two Olympus IX50 microscopes equipped with epiflouorescence and monochrome/color CCD cameras (video capable), two Leica DM IL inverted phase contrast microscopes, as well as two cell culture microscopes.

Data generated at the UPDDI is managed by a state-of-the-art network comprised of ten servers including: two Dell Poweredge (PE) R720 servers each with 128 GB memory, dual Intel Xeon 2.1GHz processors and 10 TB local storage for virtualization of workstations and servers; a PE R720 server with 16 GB memory, dual Intel Xeon 2.2GHz processors and 25 TB storage to host the GE IN Cell Miner database; a PE 2850 Oracle Database and IDBS ActivityBase® LIMS server with 8 GB memory and two Dual Core 2.8GHz CPUs; two Dell PE2950 servers with 16 & 4 GB memory and two Dual Core 2.8GHz CPUs with MS SQLServer 2008 to house the ArrayScan® VTI and ArrayScan II Cellomics Store® databases; a PE 2850 server with 8 GB memory and two Dual Core 2.8GHz CPUs with MS-SQLServer 2008 to house the Molecular Devices MDCStore database; a high-performance dual-quad core Dell PE2950 application server with four analysis engines and two high-speed dual quad-core CPUs to house the Definiens image intelligence suite; and two Dell PowerEdge (PE) 2850 Windows file servers with 8 GB memory and two Dual Core 2.8 GHz CPUs. A scalable Dell Compellent SC4020 storage array, initially configured with 30TB capacity, can be expanded to 1 PB raw capacity as needed, and a scalable EMC CX300 Storage Area Network (SAN) solution is currently the central data warehouse with 36 TB formatted storage. The UPDDI servers are hosted at UPitt’s secure, state of the art Network Operations Center with 24/7 monitoring and redundant power and cooling systems. Active data is backup up daily and static data archived semiannually.

Analysis of large and complex data sets is handled through LIMS and HTS/HCS analysis software including two IDBS ActivityBase™ LIMS with Chemistry Selection Assistants, and Protocol Transfer Assistants seats; two IDBS SARview seats; three licenses for the IN Cell Investigator HCS software; two Cellomics platinum packages of Bio-Application image analysis algorithms (V2, V3 and V4) for their ArrayScan® imaging platforms (Cell Cycle, Cell Health Profiling, Cell Motility, Cell Spreading, Compartmental Analysis AS, Cyto-Cell Membrane, Cyto-Nuc Translocation, Extended Neurite OG, GPCR Signaling, Micronucleus, Molecular Translocation, Morphology Explorer, MP Cytotoxicity, Neuronal Profiling, Neurite Outgrowth, Spot Detector, Target Activation, and Tube Formation); four vHCS™ Discovery Toolbox licenses; four vHCS™ View licenses; 6 Spotfire® data visualization licenses; MetaXpress and Acuity site licenses; a Definiens image intelligence suite for custom image analysis including one developer, one architect, and two Cellenger licenses; and the Microphysiology Systems database (MPS-Db), implemented in Django and PostgreSQL, for the design, execution and interpretation of studies in microfluidic organ models.