UCSF Helen Diller Family Comprehensive Cancer Center

CORES: Immunohistochemistry

The mandate of the Immunohistochemistry and Molecular Pathology Core is to facilitate immunohistochemical and molecular genetic analyses of tumor materials by:

  • Performing routine immunohistochemical (IHC) analyses of normal and tumor sections.
  • Performing IHC analysis of normal and tumor tissue arrays.
  • Develop and share protocols for IHC and molecular analysis.
  • Perform section-based FISH analyses for genomic copy number.
  • Collaborate with members in microdissection and extraction of DNA and RNA from tumor sections.
  • Perform genomic mapping onto metaphase chromosomes of human and mouse DNA clones.

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Services


Immunohistochemistry
Routine Immunohistochemistry Staining
Standard protocols have been developed for a number of antibodies (see list >). Sections of paraffin-embedded tissue to be stained should be brought to the Core laboratory, 2340 Sutter Street, Room S-241, between 8:00 am and 4:30 pm Monday-Friday. The tissue must be accompanied by a completed work order form. If tissue sections are not available, the paraffin block may be brought to the Core laboratory for sectioning.  Prior to the commencement of work, a recharge account needs to be set up with the Coreadmin System. Contact Maria-Elena Moncada at coreadmin@cc.ucsf.edu or 415.353.9731 to set up the account.

Protocol Development for Novel Antibodies
The laboratory assists in the development of protocols for novel antibodies. Investigators should meet with Core Manager (415.476.8825) and/or Director Dr. Rick Baehner (415.885.7301) to discuss this service. The investigator should be prepared to provide the antibody, references of its use in immunohistochemistry, positive and negative controls, information from Western analyses, and sections of the tissues to be stained.

Molecular Pathology
FISH for Copy Number
FISH analysis of specimens for copy number of specific loci, and specific translocations, on a cell-by-cell basis. This is available for cultured cells (interphase or metaphase), and for paraffin sections of formalin-fixed tissue.

FISH Mapping of DNA Probes
FISH mapping of unique sequence DNA probes to metaphase chromosomes. Metaphase spreads of human and mouse chromosomes are available through the laboratory, while other species must be prepared prior to mapping. Contact the Core for details on metaphase preparations. DNA probes should be at a concentration of 400-800 ug/ul in TE or water. 510 ugs of DNA is required.

Preparation of Metaphase Spreads for Cytogenetic Analysis
The laboratory can culture and prepare metaphase spreads if cultured cells are provided. Although the laboratory will accept slides, preferably the lab will prepare the metaphase spreads and the user need only supply the growing cell line. Cell cultures should be delivered to the lab within 24 hours after passage. Bring cell cultures to the lab no later than 12 noon Monday-Thursday. Cells can be brought to the Mt. Zion Clinical Lab to Sandy DeVries,  2340 Sutter Street, Room S341.  Phone:  476-3822.

Education

For a fee, training in IHC techniques is informal and hands-on, and takes place as time permits. Contact Loretta Chan to schedule a session. Core personnel are also available to train researchers in molecular cytogenetic techniques, comprising both didactic sessions and hands-on laboratory experience.

 


Additional Resources
A searchable database of core facilities at all UCSF campus locations, provided by the Clinical and Translational Science Institute at UCSF, is available here.