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University of Colorado Cancer Center

University of Colorado Cancer Center, A National Cancer Institute-designated Comprehensive Cancer Center
Laser Capture Microdissection | University of Colorado Cancer Center
 

Laser Capture Microdissection Service

Laser Capture Microdissection (LCM) is a technique that allows scientists to lift specifically targeted cells from a section of tissue, leaving behind unwanted cells that could contaminate the molecular purity of subsequent analysis. This technology, which was developed in the late 1990s at NIH in collaboration with bioengineering and cancer pathology groups, is precise, accurate and semi-automated.

LCM uses a laser beam to activate a special transfer film. The film bonds to cells that have been identified and targeted by microscopy. From this film, macromolecules can be isolated and studied in downstream analyses.

An infrared (IR) laser is used for single cells and small populations of cells, while an ultraviolet (UV) laser can be used for large populations of cells.  Starting tissue sections (5-7um) can be frozen or fixed/paraffin embedded, however it is ideal to use frozen sections for most downstream applications.

Why Use LCM in Gene Expression Studies?

To reveal accurate, cell-type specific expression profiles that otherwise might be obscured in mixed cell samples.

Technology isolates homogeneous populations of cells from tissue sections   

  • Cells can be collected based on morphologic or immunohistologic features
  • Machine can recognize color (stains) or fluorescence
  • Cells are harvested for DNA, RNA or Protein analysis

    • Equipment

    XT LCM Machine

    The ArcturusXT™ microdissection system is a unique combination of IR laser-enabled LCM and UV Laser Cutting in one platform. The open, modular design of the ArcturusXT™ system provides unparalleled research flexibility and versatility.

  • We will train you to use the machine, and provide technical assistance and reagents for dealing with LCM-captured material for subsequent molecular analysis.
  • The LCM microscope has brightfield (including Phase, DCIM) as well as fluorescent capabilities.
  • (We have a stage that accommodates regular microscope slides (75 mm x 25 mm) as well as large slides (75 mm x 38 mm and/or 75 mm x 50 mm).
  • The machine also has the capability to be trained by the user to recognize cells based on morphology or immunohistochemical staining. We can store customized recognition programs.
    •
    Filters Include Excitation Emission
    Red

    570-630 nm

    		 >655 nm
    Green 503-548 nm >565 nm
    Blue 455-495 nm >410 nm

     

    Agilent 2100 Bioanalyzer

    We have the capability to analyze your RNA or DNA

    RNA

  • Nanochip = 12 samples per chip, 1uL of ~300ng/uL per sample
  • Picochip = 11 samples per chip, 1uL of ~5ng/uL per sample

    • DNA 1000

  • PCR samples: For accurate determination of DNA concentration, the total DNA in sample must be between 0.1–50 ng/µl. If concentration of your particular PCR reaction is excessively high, dilute to 0.1–50 ng/µl in water.
  • Restriction digests: Final concentration of DNA should not exceed 50 ng/µl. Add EDTA and/or heat inactivate the restriction enzyme according to the manufacturer instructions. Restriction endonucleases in combination with non-chelated metal ions may degrade internal DNA markers used in assay kit.
  • 12 samples per chip, 1uL of sample
    •

    DNA 7500 & 12000 available upon request.