Liver Center Core Facilities

• Cell and Tissue Biology Core
• Liver Immunology Core
• Microscopy and Advanced Imaging Core
• Molecular Analysis Core
• Clinical Component
• Administrative Core

Cell and Tissue Biology Core

New Core Service under development:

• * Microarray technology and genomics. This service is being added on the advice of the External Advisory Board and feedback from the membership. The core is partnering with the UCSF Shared Microarray Facility (David Erle, director). Interested members may contact Dr. Maher for details.

Ongoing Services:

• Primary isolates of hepatocytes and individual non-parenchymal cell populations from rat and mouse liver.
• Instruction and consultation

Liver Immunology Core (changing to ANIMAL TECHNOLOGY CORE in June 2007)

New Core Services under development:

• Gene Transfer to the Liver.
  The core is preparing a store of quality-controlled gene-transfer reagents. It consults and assists with strategies for gene transfer including training in vector production and delivery.
• Purification and Transplantation of Cells from Liver or Bone Marrow.
  Core personnel provide training in cell purification by magnetic sorting or FACS sorting. They also train users in transplantation techniques.

Ongoing Services:

• Flow cytometry, using a BD multi-color LSR-II. The core provides training for novice users (required) and assists with interpretation of data. Users are responsible for cell isolations and labeling.
• Quantitative RTPCR. The core provides training on its ABI instrument, and it has a collection of primers including those for control reactions. Users provide custom primers and RNA extracts.
• Serum tests by COBAS autoanalyzer. AST and ALT from mouse serum are standard; bilirubin is being added. The core will run these tests or train frequent users on the instrument. For other tests, at present the cost of kits and standardization are the responsibility of the user.

Microscopy and Advanced Imaging Core

New Instruments and Capabilities:

• A complete stereology setup, including Stereo Investigator and Neurolucida software from MicroBrightField with solid modeling, virtual slice, and confocal modules
• Olympus microscope with isolation platform, 3-axis computer-controlled stage motor, and digital camera: This allows counting samples in a three dimensional space in an unbiased manner independent of cell size and shape of the tissue.
• A state-of-the-art 4 megabyte CCD camera for the Tecnai 10 electron microscope to acquire digital images at high resolution that can be sent directly to users.
• A state-of-the-art 4 megabyte CCD camera for the Tecnai 10 electron microscope to acquire digital images at high resolution that can be sent directly to the users.
• A 3-chip Hammamatsu Orca CCD camera for the Zeiss fluorescent microscope that can generate color images from different emissions of fluorescence or from histologically stained samples at high resolution and up to 9 images per second, thereby significantly improving and simplifying data acquisition.
• Arcturus Pyxis laser capture microscopy machine, for automated capture of selected cells for further analysis (e.g., gene expression, DNA sequencing).

New Core Services:

• Laser capture microdissection
• Stereology

Ongoing Services:

• Paraffin processing, sectioning, H/E staining
• Cryosectioning
• Special stains (histo-, immunohistochemistry)
• Confocal microscopy
• Fluorescence microscopy
• Live cell imaging
• Transmission EM
• ImmunoEM microscopy
• Histology interpretation

Molecular Structure Core

Ongoing Services:

• Protein identification by mass spec.
• Protein microsequencing. This can be performed on-line or off-line. The proteins can be obtained, for example, from immunoprecipitation experiments, tandem affinity purification or whole cell lysates. Proteins can be separated by 1-D or 2-D gel-electrophoresis or this step can be skipped and all the proteins in a sample digested together. In the latter instance, the peptides can then be fractionated into smaller more manageable subsets by using two dimensions of chromatography in combination. Frequently, the first dimension is cation exchange and the second dimension reversed phase chromatography.
• Proteome analysis
• Characterization of protein substitutions (glycosylation, sulfation, etc)

Clinical Component

New Core Service:

This Liver Center data and tissue bank is available for use by Center members. Interested investigators should contact Dr. Marion Peters (marion.peters@ucsf.edu).

Administration

Contact:
Monty Bissell montgomery.bissell@ucsf.edu
Jackie Maher jmaher@medsfgh.ucsf.edu

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