Laser capture microdissection (LCM) is a method for isolating specific cells of interest from microscopic regions of tissue that has been sectioned.[1] [2]
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Procedure
Under a microscope using a software interface, a thin tissue section is viewed and individual cells or clusters of cells are identified for isolation. When the cells of choice are in the center of the field of view, the operator selects the cells of interest using instrument software. Depending on the instrument used the software will either: fire a near-IR laser to activate transfer film on a cap placed on the tissue sample, fusing the film with the underlying cells of choice; or activate 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.
In addition to tissue sections, LCM can be performed on living cells, cell smears, and plant tissue.
Applications
The laser capture microdissection process does not alter or damage the morphology and chemistry of the sample collected, nor the surrounding cells. For this reason, LCM is a useful method of collecting selected cells for DNA, RNA and/or protein analyses. LCM can be performed on a variety of tissue samples including blood smears, cytologic preparations,[3] cell cultures and aliquots of solid tissue. Frozen and paraffin embedded archival tissue may also be used [4] . On formalin or alcohol fixed paraffin embedded tissues, DNA and RNA retrieval has been successful, but protein analysis is not possible (requires frozen section).[citation needed]
References
- ^ Emmert-Buck MR, Bonner RF, Smith PD, Chuaqui RF, Zhuang Z, Goldstein SR, Weiss RA, Liotta LA (1996). "Laser capture microdissection". Science 274 (5289): 998–1001. doi:10.1126/science.274.5289.998. PMID 8875945.
- ^ Espina V, Heiby M, Pierobon M, Liotta LA (2007). "Laser capture microdissection technology". Expert Rev. Mol. Diagn. 7 (5): 647–57. doi:10.1586/14737159.7.5.647. PMID 17892370.
- ^ Orba Y, Tanaka S, Nishihara H, Kawamura N, Itoh T, Shimizu M, Sawa H, Nagashima K (2003). "Application of laser capture microdissection to cytologic specimens for the detection of immunoglobulin heavy chain gene rearrangement in patients with malignant lymphoma". Cancer 99 (4): 198–204. doi:10.1002/cncr.11331. PMID 12925980.
- ^ Kihara AH, Moriscot AS, Ferreira PJ, Hamassaki DE (2005). "Protecting RNA in fixed tissue: an alternative method for LCM users". J Neurosci Methods 148 (2): 103–7. doi:10.1016/j.jneumeth.2005.04.019. PMID 16026852.
External links
- National Institutes of Health Laser Capture Microdissection Core Facility
- Yale Rice Transcriptional Atlas Project employing Laser Capture Microdissection
- Core Unit Chip Applications, Jena employing Laser Capture Microdissection
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