Cytogenetics Shared Resource

Robert B. Jenkins, M.D., Ph.D.
Director

The central focus of the Cytogenetics Shared Resource (CSR) is to provide routine cytogenetic and advanced molecular cytogenetic services on human and non-human samples. CSR services are available to Mayo Clinic Cancer Center members across all three campuses in Phoenix/Scottsdale, Ariz.; Jacksonville, Fla.; and Rochester, Minn.; and include:

• Cell culture
• Chromosome preparations
• Routine cytogenetic analysis
• Breakage studies
• Interphase and metaphase fluorescence in situ hybridization (FISH) analysis
• Multi-color FISH (MFISH) analysis
• Spectral karyotyping (SKY) analysis (human, mouse, and rat)
• Comparative genomic hybridization (CGH) analysis
• Probe labeling and development
• Microscope utilization by the user

Additionally, comparative genomic hybridization to array (CGHa) is under development.

The CSR has experience with all commercially-available FISH probes and probe strategies, including:

• Chromosome enumeration probes
• Single copy or locus-specific probes
• Translocation probes
• Multi-color FISH probes
• Whole chromosome paints
• Whole genomic paints for various species

In addition, the CSR produces homebrew probes for specific regions of interest and probe strategies (including enumeration, fusion, and break-apart probes), as well as whole genomic paints for human, mouse, pig, and other animal models .

Analysis can be performed on cells from blood or tissue samples from humans or experimental animals including cultured cells, fresh and frozen tissue, paraffin-embedded sections, and tissue microarrays. Other types of tissue that have been analyzed in the CSR include (but are not limited to) xenograft cells, hybrid cells from chimeric animal models, neural stem cells, and mouse embryonic fibroblasts .

The CSR is utilized for both conventional and unconventional studies. The CSR is open to determining novel uses for molecular cytogenetic techniques to meet the evolving needs of research studies. Investigators have used these services to:

• Characterize the karyotypes of tumors and cell lines
• Rule out interspecies contamination
• Determine the presence or absence of interspecies cells in xenograft or chimeric animal models
• Monitor established cell lines to rule out contamination or other changes during cell culture
• Enumerate copy numbers of chromosomes or cancer-related genes using "homebrew" FISH probes
• Determine patterns of gene-related gains and losses
• Map the location of DNA sequences, genes, or transgene insertions by using homebrew FISH probes, FISH, and SKY techniques in combination

Facilities, Equipment, and Staffing

The CSR provides training on molecular cytogenetic techniques and equipment as needed. Once trained, the user can utilize the CSR laboratory, reagents, and equipment. CSR technologists are also available for assistance.

The laboratory is equipped with the following equipment available for CSR Users:

• Six high-resolution fluorescent microscopes (with brightfield capabilities)
• One light microscope
• One inverted phase microscope
• Five computerized imaging systems, which capture, process, and analyze fluorescent and brightfield images
• One programmable slide processor for large slide volumes
• Four programmable hybridization ovens
• One incubator
• Three water baths
• One programmable oven

The laboratory is also equipped with the following FISH probe production equipment:

• One shaking incubator
• One high performance centrifuge
• One micro centrifuge
• One refrigerated water bath
• One thermal cycler
• One ultrasonic processor (Sonicator)

Cytogenetics Staff

• Director: Robert Jenkins M.D., Ph.D.
• Manager: Kari Hafner
• One FISH technologist
• One FISH probe production technologist