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Expression microdissection adapted to commercial laser dissection instruments

2011; Nature Portfolio; Volume: 6; Issue: 4 Linguagem: Inglês

10.1038/nprot.2010.202

1754-2189

Jeffrey C. Hanson, Michael A. Tangrea, Skye Kim, Michael D. Armani, Thomas J. Pohida, Robert F. Bonner, Jaime Rodriguez‐Canales, Michael R. Emmert‐Buck,

Cancer Cells and Metastasis

Laser-based microdissection facilitates the isolation of specific cell populations from clinical or animal model tissue specimens for molecular analysis. Expression microdissection (xMD) is a second-generation technology that offers considerable advantages in dissection capabilities; however, until recently the method has not been accessible to investigators. This protocol describes the adaptation of xMD to commonly used laser microdissection instruments and to a commercially available handheld laser device in order to make the technique widely available to the biomedical research community. The method improves dissection speed for many applications by using a targeting probe for cell procurement in place of an operator-based, cell-by-cell selection process. Moreover, xMD can provide improved dissection precision because of the unique characteristics of film activation. The time to complete the protocol is highly dependent on the target cell population and the number of cells needed for subsequent molecular analysis.