Title

Laser microdissection separation of pure spermatozoa from epithelial cells for short tandem repeat analysis

Authors

Authors

C. T. Sanders; N. Sanchez; J. Ballantyne;D. A. Peterson

Comments

Authors: contact us about adding a copy of your work at STARS@ucf.edu

Abbreviated Journal Title

J. Forensic Sci.

Keywords

forensic science; DNA typing; DNA mixtures; cell separation; histology; DNA isolation; SEXUAL ASSAULT EVIDENCE; CAPTURE MICRODISSECTION; TISSUE-SECTIONS; DNA; SPERM; AMPLIFICATION; STAINS; Medicine, Legal

Abstract

Short tandem repeat (STR) analysis is a valuable tool in identifying the source of biological stains, particularly from the investigation of sexual assault crimes. Difficulties in analysis arise primarily in the interpretation of mixed genotypes when cell separation of the sexual assailant's sperm from the victim's cells is incomplete. The forensic community continues to seek improvements in cell separation methods from mixtures for DNA typing. The feasibility of applying laser microdissection (LMD) technology to precisely separate sexual assault cell mixtures by visual inspection coupled with laser dissection was assessed through three experiments. First, various histological stains were evaluated for use with LMD and DNA analysis. Second, different DNA isolation methods were evaluated on LMD-collected cells. Finally, STR analysis was performed on LMD-separated sperm cells from mixtures of semen and female buccal epithelial cells. The results indicated (a) hematoxylin/eosin staining performed best in its ability to differentiate sperm and epithelial cells while exhibiting the least negative effect on further downstream analysis; (b) both QIAamp (R) and Lyse-N-Go (TM) methods were useful for recovery of DNA from LMD-collected sperm cells; and (c) LMD separation provided clear STR profiles of the male donor with the absence of any additional alleles from the female donor. This report describes an efficient, low-manipulation LMD method for the efficient separation of spermatozoa from two-donor sperm/epithelial cell mixtures.

Journal Title

Journal of Forensic Sciences

Volume

51

Issue/Number

4

Publication Date

1-1-2006

Document Type

Article; Proceedings Paper

Language

English

First Page

748

Last Page

757

WOS Identifier

WOS:000238790200005

ISSN

0022-1198

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