Tissue and Cell Molecular Analysis (TACMA) Shared Resource

The Tissue and Cell Molecular Analysis (TACMA) Shared Resource of Mayo Clinic Cancer Center is a resource of expertise, collaborative support and service for research and clinical investigators. It provides research service for pathology, immunohistochemistry, in situ hybridization, digital image analysis, laser capture microdissection, and preparation of tissues for subsequent protein extraction for members of Mayo Clinic Cancer Center. Several areas of service are being expanded and developed in the TACMA Shared Resource, one of them being incorporated as part of the Advanced Genomics Technology Center (AGTC).

The AGTC is supporting development of the Research Laboratory Information Management System (RLIMS). A proposed four-phase program, RLIMS is laying the groundwork to develop an information management infrastructure for the Mayo research laboratory environment, including Mayo Clinic Cancer Center. The vision of the RLIMS program is to develop standardized tools for research laboratory operations involving the use of human and animal biospecimens and information management across all three Mayo sites, in Scottsdale/Phoenix, Ariz.; Jacksonville, Fla.; and Rochester, Minn. This initiative enables Mayo investigators to leverage the extraordinarily rich dataset associated with specimens processed and analyzed in all the research laboratories, ensure compliance with regulatory guidelines, encourage collaboration between Mayo investigators, and improve the productivity of the Mayo research laboratories.

TACMA provides a number of other services relating to molecular analysis, including:

• Immunohistochemistry
  TACMA offers both manual and automated immunostaining using a variety of antibody detection chemistries, including DAB, AEC, and NovaRed for use with brightfield microscopy, and secondary antibodies conjugated to fluorophores for epifluorescence microscopy. The laboratory also provides customized optimization and titration of new antibodies for investigators.
• Digital Imaging
  Featuring some of the newest virtual microscopy and scanning technologies, the core offers digital scanning and detailed analysis of both brightfield and fluorescent tissue slides. Individual TMA cores and whole tissue sections can be digitally scanned and archived. These images can then be reviewed by research pathologists and other investigators.
• Tissue Handling
  With access to both a paraffin embedding station and a Histobath cryobath, TACMA has the capability of snap-freezing fresh tissues and re-embedding old, archival formalin-fixed paraffin embedded (FFPE) samples. Researchers can also have frozen samples physically dissected to enrich the percentage of benign or diseased tissue within any given block.
• Sectioning
  Having multiple microtome cutting stations and two cryostats, TACMA is equipped to handle the sectioning of both frozen and FFPE tissues. These sections can be mounted on a variety of slides, or researchers have the option of requesting such sections be placed in 1 to 2 milliliter tubes for subsequent RNA, DNA, or protein extraction. Sections from both whole blocks and tissue microarrays can be generated.
• Laser Capture Microdissection
  Utilizing the Veritas Microdissection Instrument, investigators now can isolate individual cells or groups of cells from frozen and FFPE tissue sections. Slides to be microdissected can be either brought by researchers themselves, or be prepared by TACMA at the investigator’s request. Training on TACMA’s Veritas instrument is available.
• Tissue Microarray Construction
  TACMA personnel are experienced in constructing custom tissue arrays for investigators who provide the appropriate paraffin blocks. With the acquisition of a Beacher ATA-27, the lab can now create arrays containing as many as 540 tissue cores, with each core typically being 0.6 millimeters in diameter. Smaller arrays can also be created using the Beecher instrument.

Equipment, Facilities, and Staffing

Current instruments available in the TACMA lab include the following:

• One Beecher ATA-27 automated tissue microarray constructor
• Two Bliss Digital Imaging Systems (Bacus Laboratories) for digital image capture and analysis
• One Nikon COOLSCOPE to analyze Feuglen-stained tissues
• One Hamamatsu NanoZoomer Digital Pathology (NDP) instrument for rapid digital scanning of large batches of slides
• One HistoRx PM-2000 Image Analyzer to aid in protein analysis under fluorescent conditions
• One Shandon histobath refrigerated snap-freezing tissue bath
• Two Leica CM1850 cryostats for cutting frozen sections
• One Leica RM2025 and three Leica RM2255 rotary microtomes for cutting FFPE sections
• One Leica IP S slide printer
• One Leica CV5030 automated coverslipper
• One Veritas Laser Capture Microdissection system from Molecular Devices/Arcturus
• One Olympus BX50 microscope with a multihead attachment
• Three automated immunohistochemical stainers from the Autostainer series of instruments produced by the Dako Corporation
• One Sanyo Ultra-Low MDF-U73VC VIP Series -80°C freezer for storage of frozen project specimens and slides

TACMA Team Leadership

Director: Wilma Lingle, Ph.D., 507-538-1287
Co-Director: Ricardo Lloyd, M.D., Ph.D., 507-284-4022
Supervisor: Jason Carnahan 507-538-0709
Asst. Supervisor: Karla Kopp 507-266-5115

Additional team members include a laser microdissection specialist, a research technologist who specializes in immunohistochemistry, an image analysis specialist and eight other researchers and assistants.