Digoxigenin labeling and laser capture microdissection of male cells

Institute of Legal Medicine, Ludwig-Maximilians-University of Munich, Frauenlobstrasse 7a, 80337 Munich, Germany.
Deutsche Zeitschrift für die Gesamte Gerichtliche Medizin (Impact Factor: 2.71). 11/2005; 119(6):374-7. DOI: 10.1007/s00414-005-0523-2
Source: PubMed


Laser capture microdissection (LMD) is a relatively new technique for the isolation of single cells. The application in forensic investigations has become more and more widespread, especially to select spermatozoa out of mixtures with vaginal cells. In particular in cases with low numbers of sperm it could be profitable to isolate all male cells (e.g. sperm and male epithelial cells) instead of focussing on the sperm only. Therefore, the specific labelling and detection of the male cells in a male/female cell mixture is necessary. In order to label all cells carrying a Y-chromosome we used a digoxigenin labelled chromosome Y hybridisation probe (Q Biogen). The stained cells were isolated with the SL microCut LMD system from Molecular Machines & Industries AG (MMI). At least ten diploid male cells were required to obtain a partial STR profile, with 20 cells, a full profile could be obtained.

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    • "Fluorescence in situ hybridisation (FISH) probes, specific for X and Y chromosomes, are used to distinguish male from female cells. This application becomes widespread in forensic investigations of sexual assault cases with low numbers of sperm cells or male/female mixtures of blood or epithelial cells (Anslinger et al. 2005; Vandewoestyne and Deforce 2010). In medical sciences, LCM is used in cancer research to determine the cell-specific gene expression profile from heterogeneous human tissues (Curran et al. 2000; Cho-Vega et al. 2005). "
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    • "The different cell types can be recognized by morphological characteristics, various chemical staining or fluorescence labeling techniques. These methods allow to establish a clear DNA profile from few cells present in a mixture samples that otherwise had not be detected while swabbed by the major component and not detectable in the profile ( Elliott et al., 2003; Anslinger et al., 2005; Anoruo et al., 2007 ; Sanders et al., 2006). With laser micro dissection techniques ( Anslinger et al., 2007; Vandewoestyne et al., 2009), it has been shown that cells derived from a male contributor can be analyzed separately from those derived from a female contributor after morphological or fluorescent labeling identification. "
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    • "Similarly, specific detection and isolation of male cells in male/female mixtures such as fingernail scrapings, bite or licking traces and male/female blood mixtures could be a major advantage. Therefore, several groups developed staining methods to perform sex-specific labelling of cells for LCM [37–41]. To distinguish male from female cells, fluorescence in situ hybridization (FISH) is performed using Y chromosome specific probes. "
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