Simon C. Watkins Ph.D. FRC Path.
Professor, Cell Biology and Physiology and Immunology
Director Graduate Program in Cell Biology and Physiology
Vice Chair, Cell Biology and Physiology
Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St .
Pittsburgh PA 15261
Tel:412-648-3051
Fax:412-648-2797

Currently available or expected date of availability

Currently available

Requirements for use (by collaboration or fee-for-service, application required, etc.)

Collaboration and if successful and useful co-submission of Grants

Brief description of services or resources (a paragraph or two)

The CBI is one of the largest optical imaging centers in the country It is housed in the medical research facility of the University of Pittsburgh Medical School in approximately 5,500 sq ft. of space. This space has been designed as a dedicated, state of the art imaging center, and has fully equipped microscopy suites, computer labs, and wet and dry bench space for light and electron microscopic preparations.

Apart from office space for the Director, the Assistant Director and post-doctoral fellows, desk areas are provided for the 10 full-time research specialists who work within the facility. Importantly, there is sufficient undedicated bench space within the facility for users to conduct several concurrent projects.

CBI Core equipment

Microscopes:

1 Leica TCS NT Confocal Microscope (Inverted)
1 Leica TCS-SL Confocal Microscope (Upright)
2 Olympus Fluoview 1000 Confocal Microscope (Inverted)
1 Olympus F500 Confocal Microscope (Upright)
1 Zeiss 510 Meta Confocal Microscope(Inverted)
1 Perkin Elmer Spinning Disk Confocal Microscope
1 Olympus DSU Spinning Disk Confocal Microscope
1 Noran OZ (retrofitted to PC Platform) Confocal Microscope
Optiscan Confocal Endoscope/Microscope
1 Olympus FluoView/Coherent Mira Multiphoton Microscope
3 Multimode microscopes ((Automated XY, Ratioing, FRET) (one with microinjection capabilities)
2 Multimode Widefield/TIRF/ microscopes (one with microinjection capabilities)
1 In vivo murine imaging system (luciferase imaging)
1 Olympus Multimode (3D, 2D fluorescence, high speed) dissecting microscope
1 Olympus BX51 light microscope (Brightfield, darkfield, epifluorescence DIC)
2 Olympus Provis light microscope (Brightfield, darkfield, epifluorescence DIC)
1 Nikon Eclipse 800 microscope (Brightfield, darkfield, epifluorescence DIC)
1 JEOL 1011CX Transmission Electron microscope
1 JEOL 9335 Field Emission Gun SEM
1 JEOL 1210 Transmission Electron microscope

Microtomes:

3 Microm cryostats
3 Reichert Ultracut (R,S,E Ultramicrotomes 2 with cryokits)

Image analysis:

2 Scanners (flat bed and slide)
23 Pentium based PCs all with MetaMorph, and Photoshop

Services available within the CBI

1. Light Microscopic Services

Light microscopy using a variety of standard histological or optical contrasting methods such as differential interference contrast (DIC) or phase contrast microscopy are continually provided as a service to PROGRAM members. The Core also provides a full spectrum of fluorescent microscopic methods. Three major techniques, all of which allow multicolor analyses of tissues and cells, are employed.

Confocal and 2 Photon Laser Scanning Microscopy (CLSM)
This technique, which "optically sections" material, allows visualization of fluorochromes within tissues and cells with exquisite spatial resolution. In addition, quantitative measurements of fluorescence intensity may be made, and serial sections may be rendered in three dimensions allowing determination of spatial distributions of cellular label. This technology is already used extensively by Pittsburgh Cancer researchers. Currently the imaging core provides access to 8 confocal systems and a single 2 photon system.
Fluorescence Microscopy and Image Analyses.
Using standard fluorescence or brightfield microscopy, highly sensitive color camera systems and computer image processing, we have provided several research groups with access to qualitative and quantitative microscopic imaging tools. These projects have generally focused on assessment pathology in sectioned material. Quantitatively, we have investigated a wide variety of parameters tumor growth, immune cell infiltration, reduction in tumor size etc. At the present time, the core provides access to 4 such instruments. In each case, the instruments are equipped with fluorescence and DIC imaging capabilities, and all image collection is performed using high resolution, cooled CCD cameras.
Multicolor Fluorescence Microscopy, Live Cell Microscopy, and Ratio imaging.
Live cell imaging is expected to become an essential service provided by CBI to the PROGRAM members. This technology when combined with multimode, multiparameter fluorescence imaging allows molecular events to be studied in living cells in situ . (for example see the cell migration assays described below/. At the present time the core has 5 instruments available to researchers. Apart from live cell imaging, these instruments have been expanded to allow confocal imaging (3), micro injection (2) and TIRF imaging (1). We expect that given the potentially complex 3D nature of the samples we will image in this proposal, live cell Confocal will play and increasingly important role in this project.

2. Electron Microscopy Services.

The Core is equipped with both transmission and scanning electron microscopes as well as all necessary support equipment. The services provided include: standard transmission electron microscopy; scanning electron microscopy; thin frozen section immuno-electron microscopy; electron microscopic autoradiography; and freeze fracture electron microscopy. These services are commonly used as an essential extension of the light microscopy services, for example to localize an antigen with higher resolution than is available by light microscopy or to provide greater structural resolution of cells and tissues or to provide structural studies of relevant subcellular species .

3. Computer-Aided Morphometry and Image Analysis

An important facet of the services provided by the Core is the design and implementation of image processing and morphometric analysis algorithms. These analyses are generally performed on 2, 3, or 4 dimensional digital data sets collected using the microscopes within the Core. Commonly, the Core staff will develop algorithms for feature extraction and quantitation following discussions with the biostatistics core. This ensures optimal experimental design and statistical utility. Currently, the Core has over 20 work stations available to investigators, with over 24 Terabytes of online storage. This facet of core will be invaluable to numerous investigators within the program.