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Core Vision Research at the Eye and Ear Institute See the Difference The Department of Ophthalmology at the University of Pittsburgh’s Eye and Ear Institute has long held a prestigious competitive Core Facility grant award from the National Eye Institute. This funding has supported our vision researchers with specialized expertise and equipment for over two decades, and contributes substantially to the Department’s reputation as a world class leader in vision research. What does the Core offer? The Core has five modules offering expertise and access to resources for different areas of research needs: (1) Imaging Acquisition & Analysis, (2) Tissue Culture & Histology, (3) Genomics, Proteomics & Molecular Biology, (4) Flow Cytometry & Cell Sorting and (5) Custom Fabrication. Each module is staffed by a full-time dedicated research specialist, called the Module Supervisor, and it is overseen by a faculty-level Module Director (see reverse for details). Can I use the Core? The Core is available to all eligible Participating Investigators. A researcher qualifies as a Participating Investigator if they work at the University of Pittsburgh and are studying vision research. Others may utilize the Core's services only if they partner in a collaboration with a qualifying Participating Investigator who contributes funding, not merely consultation or advisory support. Who gets priority for Core time and resources? Since many investigators use the Core, the time that the Core personnel can devote to individual requests is limited, and specialized equipment must be shared. In general, usage is given to approved projects on a firstcome-first-served basis. Among eligible users, those with R01 funding from the National Eye Institute receive priority over those without. How do the Module Supervisors provide support? All modules provide personnel training for equipment use and technical skill transfer, and all will consult on experimental design and data analyses. Certain modules will perform the benchwork itself on your behalf, time and resources permitting. Such benchwork is negotiated on a project-to-project basis. How do I get my project approved? The Module Supervisors generally plan their own work schedules, so you can communicate your needs directly to them. In order to receive the Director’s approval, you will need to provide the Supervisor with information about the vision-related grant that funds this aspect of your project, and any associated institutional protocol numbers (e.g. IACUC, IBC, DNA/RNA usage, etc.). What do I need to pay for? Labor is provided at no cost to the Participating Investigator. This includes time the Core Supervisor spends doing benchwork, evaluating data, and training personnel. Supplies, however, are often the responsibility of the Participating Investigator, and the policies for individual modules vary. What are my responsibilities as a Core user? You must agree to acknowledge your utilization of the Core by citing the core grant for vision research -P30 EY08098 - on your posters, in your talks, and in your manuscripts. Also, the individual modules each have rules for the proper use and care of Core equipment, which you will be expected to follow. Image Acquisition & Analysis Optical Coherence Tomography (OCT) Whole-mount, section, and live-cell microscopy Digital imaging for stationary specimens Phase, Darkfield, and Light field systems Confocal and fluorescent microscopy systems Automated real-time imaging and micromanipulation Time-lapse and z-stacks over multiple stage positions Software training for Metamorph, Image-J, and Photoshop Writing software for specialized analyses Microscopy theory and equipment training Troubleshooting problems Gel imaging systems CO-DIRECTORS DIRECTOR Kira Lathrop, MAMS 1026 Eye & Ear Institute (412) 647-3492 [email protected] Dr. Ian A. Sigal, PhD 930 Eye & Ear Institute (412) 864-2220 [email protected] Dr. Larry Kagemann, PhD 834 Eye & Ear Institute (412) 648-6409 [email protected] Tissue Culture & Histology Immortalized cell culture Primary cell culture from ocular tissues Hybridoma growth and supernatant harvest Human embryonic stem cells Virus amplification, purification, and titering Cell banking Formalin fixation Cryostat embedding and sectioning (8-30um) Paraffin embedding and sectioning (5-10 um) Immunohistochemistry H&E and other counter stains SUPERVISOR DIRECTOR Katherine Davoli 928 Eye & Ear Institute (412) 647-8256 [email protected] Dr. James L. Funderburgh, PhD 1013 Eye & Ear Institute (412) 647-3853 [email protected] Genomics, Proteomics & Molecular Biology Realtime and quantitative PCR* Gene chip arrays* Prep and analysis of DNA and RNA* Preparation and screening of phage libraries† Generation of subtractive libraries† DNA mutagenesis and expression† DNA-protein, and protein-protein interactions† RNA mapping and accurate quantification† Virus vector design and derivation† Immunoprecipitations† 2D and 3D Western blots† *SUPERVISOR †DIRECTOR Dr. Stephen Harvey, PhD 1014 Eye & Ear Institute (412) 647-8919 [email protected] Dr. Paul R. Kinchington PhD 1016 Eye & Ear Institute (412) 647-6319 [email protected] SUPERVISOR DIRECTOR Nancy Zurowski 1048 Eye & Ear Institute (412) 647-1413 [email protected] Dr. Robert L. Hendricks, PhD 922 Eye & Ear Institute (412) 647-5754 [email protected] Flow Cytometry & Cell Sorting Cytometer equipment maintenance Personnel training Fluorochrome selection consultation Sample fixation Gating selection for optimal results Live and fixed cell sorting Plot creation and data analysis Statistical validation for side populations Custom Fabrication Computer-Aided Design (CAD) 3D printing Prototyping 3D scanning/digitizing Custom metalwork and machining Laser-cutting Access to Swanson Center in Bioengineering DIRECTOR Dr. Matthew A. Smith, PhD 914 Eye & Ear Institute (412) 647-2313 [email protected]