Laser capture microdissection

Laser capture microdissection (LCM) is a method for isolating pure cells of interest from specific microscopic regions of tissue sections.cite journal |author=Emmert-Buck MR, Bonner RF, Smith PD, Chuaqui RF, Zhuang Z, Goldstein SR, Weiss RA, Liotta LA |title=Laser capture microdissection |journal=Science |volume=274 |issue=5289 |pages=998–1001 |year=1996 |pmid=8875945 |doi=10.1126/science.274.5289.998] cite journal |author=Espina V, Heiby M, Pierobon M, Liotta LA |title=Laser capture microdissection technology |journal=Expert Rev. Mol. Diagn. |volume=7 |issue=5 |pages=647–57 |year=2007 |pmid=17892370 |doi=10.1586/14737159.7.5.647]

Procedure

A transparent transfer film is applied to the surface of a tissue section. Under a microscope, the thin tissue section is viewed through the glass slide on which it is mounted and microscopic clusters of cells are selected for isolation. When the cells of choice are in the center of the field of view, the operator pushes a button which activates a near IR laser diode integral with the microscope optics. The pulsed laser beam activates a precise spot on the transfer film, fusing the film with the underlying cells of choice. The transfer film with the bonded cells is then lifted off the thin tissue section, leaving all unwanted cells behind.

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,cite journal |author=Orba Y, Tanaka S, Nishihara H, Kawamura N, Itoh T, Shimizu M, Sawa H, Nagashima K |title=Application of laser capture microdissection to cytologic specimens for the detection of immunoglobulin heavy chain gene rearrangement in patients with malignant lymphoma |journal=Cancer |volume=99 |issue=4 |pages=198–204 |year=2003 |pmid=12925980 |doi=10.1002/cncr.11331] cell cultures and aliquots of solid tissue. Frozen and paraffin embedded archival tissue may also be used cite journal |author=Kihara AH, Moriscot AS, Ferreira PJ, Hamassaki DE |title=Protecting RNA in fixed tissue: an alternative method for LCM users |journal=J Neurosci Methods |volume=148 |issue=2 |pages=103–7 |year=2005 |pmid=16026852 |doi=10.1016/j.jneumeth.2005.04.019] . On formalin or alcohol fixed paraffin embedded tissues, DNA and RNA retrieval has been successful, but protein analysis is not possible (requires frozen section).Fact|date=April 2008

References

External links

* [http://home.ccr.cancer.gov/LOP/Research/lcm/Default.asp National Institutes of Health Laser Capture Microdissection Core Facility]
* [http://bioinformatics.med.yale.edu/rc/overview.jspx Yale Rice Transcriptional Atlas Project] employing Laser Capture Microdissection
* [http://www.med.uni-jena.de/chip/English/titelundtexteng.htm Core Unit Chip Applications, Jena] employing Laser Capture Microdissection
* [http://www.leica-microsystems.com/lmd6000 Leica Microsystems] commercial non-contact Laser Microdissection and Imaging Workstations