page_title: Laser capture microdissection
section_title: Procedure
hierarchical_section_title: Laser capture microdissection / Procedure
caption_attribution_description: English: Laser Capture Microdissection
context_page_description: Laser capture microdissection, also called microdissection, laser microdissection, or laser-assisted microdissection, is a method for isolating specific cells of interest from microscopic regions of tissue/cells/organisms.
context_section_description: Under a microscope using a software interface, a tissue section (typically 5-50 micrometres thick) is viewed and individual cells or clusters of cells are identified either manually or in semi-automated or more fully automated ways allowing the imaging and then automatic selection of targets for isolation.  Currently six primary isolation/collection technologies exist using a microscope and device for cell isolation.  Four of these typically use an ultraviolet pulsed laser (355 nm) for the cutting of the tissues directly or the membranes/film, and sometimes in combination with an IR laser responsible for heating/melting a sticky polymer for cellular adhesion and isolation. IR laser provides a more gentle approach to microdissection. A fifth ultraviolet laser based technology uses special slides coated with an energy transfer coating which, when activated by the laser pulse, propels the tissue or cells into a collection cap.
  The laser cutting width is usually less than 1 μm, thus the target cells are not affected by the laser beam. Even live cells are not damaged by the laser cutting and are viable after cutting for cloning and reculturing as appropriate.
  The various technologies differ in the collection process, possible imaging methods (fluorescence microscopy/bright field microscopy/differential interference contrast microscopy/phase contrast microscopy/ etc.) and the types of holders and tissue preparation needed before the imaging and isolation.  Most are primarily dedicated micro-dissection systems, and some can be used as research microscopes as well, only one technology (#2 here, Leica) uses an upright microscope, limiting some of the sample handling capabilities somewhat, especially for live cell work.
  The first technology (used by Carl Zeiss PALM) cuts around the sample then collects it by a "catapulting" technology.  The sample can be catapulted from a slide or special culture dish by a defocused U.V laser pulse which generates a photonic force to propel the material off the slide/dish, a technique sometimes called Laser Micro-dissection Pressure Catapulting (LMPC). The dissected material is sent upward (up to several millimetres) to a microfuge tube cap or other collector which contains either a buffer or a specialized tacky material in the tube cap that the tissue will adhere to.  This active catapulting process avoids some of the static problems when using membrane-coated slides.
  Another process follows gravity-assisted microdissection method that turns on gravity to collect samples in tube cap under the slide used (used by ION LMD system, Jungwoo F&B). In case of this system, it moves the motorized stage to cut the cells of interests, keeping the laser beam fixed. And the system uses a 355 nm Solid-state laser(UV-A) which is the safest way to cut the tissues without RNA or DNA damage.
  Another closely related LCM process (used by Leica) cuts the sample from above and the sample drops via gravity (gravity-assisted microdissection) into a capture device below the sample. The different point with upper one is, the laser beam here is moving to cut tissue by moving dichroic mirror.
  When the cells (on a slide or special culture dish) of choice are in the center of the field of view, the operator selects the cells of interest using instrument software. The area to be isolated when a near-IR laser to activate transfer film on a cap placed on the tissue sample, melting the adhesive which then fuses the film with the underlying cells of choice (see Arcturus systems); and/or by activating a UV laser to cut out the cell of interest. The cells are then lifted off the thin tissue section, leaving all unwanted cells behind. The cells of interest are then viewed and documented prior to extraction.
  The fourth UV based technology (used by Molecular Machines and Industries AG) offers a slight difference to  the 3rd technology here by essentially creating a sandwich of sorts with slide>sample>and membrane overlying the sample by the use of a frame slide whose membrane surface is cut by the laser and ultimately picked up from ab
caption: Laser Capture Microdissection
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