Laser capture microdissection

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Laser capture microdissection transfer of pure breast duct epithelial cells. Left panel shows tissue section with selected cells removed. Right panel shows isolated epithelial cells on transfer film.

Laser Capture Microdissection (LCM) is a method for isolating pure cells of interest from specific microscopic regions of tissue sections.^[1] ^[2]

1 Procedure
2 Applications
3 References
4 External links

[edit] 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.

[edit] 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]}

[edit] 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. 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.