Trends in Genetics
Volume 14, Issue 7, 1 July 1998, Pages 272-276
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Laser-capture microdissection: opening the microscopic frontier to molecular analysis

https://doi.org/10.1016/S0168-9525(98)01489-9Get rights and content

Abstract

As the list of expressed human genes expands, a major scientific challenge is to understand the molecular events that drive normal tissue morphogenesis and the evolution of pathological lesions in actual tissue. Laser capture microdissection (LCM) has been developed to provide a reliable method to procure pure populations of cells from specific microscopic regions of tissue sections, in one step, under direct visualization. The cells of interest are transferred to a polymer film that is activated by laser pulses. The exact morphology of the procured cells (with intact DNA, RNA and proteins) is retained and held on the transfer film. With the advent of LCM, cDNA libraries can be developed from pure cells obtained directly from stained tissue, and microhybridization arrays of thousands of genes can now be used to examine gene expression in microdissected human tissue biopsies. The fluctuation of expressed genes or alterations in the cellular DNA that correlate with a particular disease stage can ultimately be compared within or between individual patients. Such a fingerprint of gene-expression patterns can provide crucial clues for etiology and might, ultimately, contribute to diagnostic decisions and therapies tailored to the individual patient. Molecules found to be associated with a defined pathological lesion might serve as imaging or therapeutic targets.

Section snippets

History

Preparative techniques to isolate organelles and cells for subsequent analysis of their macromolecular and biochemical contents have long been critical in cell and molecular biology. Modern techniques, such as flow cytometry with cell sorting and affinity-labeled magnetic beads, allow separation of subpopulations from heterogeneous pools of single cells in suspension. To apply these techniques to tissues, there is a requirement for the dissolution of intercellular adhesion and the formation of

LCM

LCM provides a method for the reliable procurement of specific cells. A slide with sectioned tissue (e.g. 5 μm thick) is placed on the stage of the microscope (Fig. 1) and an area of interest is selected by the user. The transport arm then places a transparent ethylene vinyl acetate (EVA) thermoplastic film in precise apposition to the tissue specimen. A joystick is used to move the slide easily until the cell(s) of interest are in the center of the optical field. With the simple push of a

Applications and the future of LCM

With the ease of procuring a homogeneous population of cells from a complex tissue using the LCM, opportunities for the molecular analysis of pathological processes are significantly enhanced. LCM can be used to capture any specific phenotypes identifiable by optical microscopy within a tissue sample. Cellular function within complex organs can now be analyzed by separation of adjacent, interacting and morphologically identifiable subunits, which can be analyzed separately by two-dimensional

References (15)

  • C. Mies

    Hum. Pathol.

    (1994)
  • R.F. Chuaqui

    Urology

    (1997)
  • R.F. Bonner

    Science

    (1997)
  • M.R. Emmert-Buck

    Science

    (1996)
  • J. Going

    J. Pathol.

    (1996)
  • D. Shibata

    Am. J. Pathol.

    (1992)
  • ...
There are more references available in the full text version of this article.

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