Trends in Genetics
Laser-capture microdissection: opening the microscopic frontier to molecular analysis
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
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